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Laidlaw Waste Systems Jonathan Greenberg. Examples include wastes from the production of chemicals, iron and steel, and food goods. Erode of Pacific Environmental Services, Inc. Atkinson as Work Assignment Manager. The enhancements described in this Addendum include changes to the processing of multi-year averages for post PM10 NAAQS analyses, enhancements to the model which were formerly available in draft form as ISCST datedenhancements to the model for air toxics applications, and an option to specify variable emission rate factors that vary by season, hour-of-day, and day-of-week.
User instructions for these enhancements are provided below. This guidance utilizes the expected second high value of the hour NAAQS replaced by a 3-year average of the 99th percentile value of the frequency distribution and a 3-year average of the annual mean. Since the Guideline on Air Quality Modeling precludes the use of a 3-year data set, a policy was established that uses unbiased estimates of the 3-year averages, utilizing all meteorological data both single and multiple years of data available.
An unbiased estimate of the 99th percentile is the fourth highest concentration, if one year of meteorological data are input to the model, or the multi-year average of the fourth highest concentrations, if more than one year of meteorological data are input to the model. Similarly, an unbiased estimate of the 3-year average annual mean is simply the annual mean, if only one year of meteorological data are input to the model, or the multi-year average annual mean if multiple years of meteorological data are used.
Analogously to the original NAAQS situation, the entire area is in compliance when the highest fourth high or highest average fourth high and the highest annual mean or the highest average annual mean are less than or equal to the NAAQS.
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In this case, the model will compute an average of the fourth highest concentrations at each receptor across the number of years of INDEX-1 meteorological data being processed. For a single year of data, the model will report the fourth highest concentration at each receptor. For a five year period of data, the model will report the average of the five fourth-highest values at each receptor.
Also, for multiple year data files, the annual average will first be calculated for each individual year of data, and the average of these across the number of years will be calculated.
This processing of the annual average across multiple years may give slightly different results than the PERIOD average across the same time period, due to differences in the number of calms from year to year. Use of the PERIOD average or a short-term average other than hour will result in a fatal error message being generated. The model will only process complete years of meteorological data, although there is no restriction on the start date for the data.
If less than one complete year of data is processed, a fatal error message will be generated. If additional meteorological data remains after the end of the last complete year of data, the remaining data will be ignored, and a non-fatal warning message will be generated specifying the number of hours ignored.
Multiple year analyses should be accomplished by including the multiple years of meteorology in a single data file. Since the hour average design values for post PM10 analyses may consist of averages over a multi-year period, they are incompatible with the EVENT processor. The syntax and type are now as follows: Optional, Non-repeatable where H6H is a new secondary keyword that identifies this as a pre analysis, the Savfil parameter specifies the filename for saving the results arrays at the end of each year of processing, and the Inifil parameter specifies the filename to use for initializing the results arrays at the beginning of the current year.
The Inifil parameter is optional, and should be left blank for the first year in the multi-year series of runs. Memory Allocation The revised ISCST3 model will allocate data storage as needed based on the number of sources, receptors, source groups, and other input requirements, up to the maximum amount of memory available on the computer being used.
The minimum system requirements for this version of the model are a or higher processor with a math coprocessor and at least 2 MB of extended memory.
The ISCST3 model preprocesses the model runstream input file to determine the data storage requirements for a particular model run, and then allocates the input data arrays before processing the setup data.
Once the setup processing is completed, the model allocates storage for the result arrays. If the allocation is unsuccessful, then an error message is generated by the model and further processing is prevented. Also, an estimate of the total amount of memory needed for a particular run is printed out as part of the first page of printed output.
The parameters that are established at model runtime are as follows: Other parameters are set to the actual numbers required for a particular model run. A change has also been made that affects the length of filenames that may be specified in the ISCST3 model input file.
FOR, which is initially assigned a value of This PARAMETER is now used to specify the maximum length of individual fields on the input runstream image, and also to declare the length of all filename and format variables. This includes the input and output filenames specified on the command line. The data in the included file will be processed as though it were part of the runstream file.
The contents of the included file must be valid runstream images for the applicable pathway. If an error is generated during processing of the included file, the error message will report the line number of the included file.
All three of the area source types use the same numerical integration algorithm for estimating impacts from area sources, and are merely different options for specifying the shape of the area source.
The AREA source keyword may be used to specify a rectangular-shaped area source with arbitrary orientation; the AREAPOLY source keyword may be used to specify an area source as an irregularly-shaped polygon of up to 20 sides; and the AREACIRC source keyword may be used to specify a circular-shaped area source modeled as an equal-area polygon of up to 20 sides.
Note that the source elevation, Zs, is an optional parameter. The source coordinates may be input as Universal Transverse Mercator UTM coordinates, or may be referenced to a user-defined origin. This source type option provides the user with considerable flexibility for specifying the shape of an area source. The number of vertices or sides used to define the area source polygon may vary between 3 and The remaining vertices may be defined in either a clockwise or counter-clockwise order from the point used for defining the source location.
The model will automatically generate a regular polygon of up to 20 sides to approximate the circular area source.
The polygon will have the same area as that specified for the circle. The approach used by the SCEVI option is to sample the meteorological data at a user-specified regular interval to approximate the long-term i.
Since wet deposition does not occur at regular intervals, the user can also specify a separate wet sampling interval to reduce the uncertainty introduced by sampling for wet deposition. Therefore, the annualized deposition rates for the two types of deposition are calculated separately. Studies have shown that the uncertainty in modeled results introduced by use of the SCEVI option is generally lower for area sources than for point sources.
When only the regular sampling is selected, all hourly impacts concentration, dry deposition flux and the wet deposition flux are calculated in the normal fashion for each sampled hour. The annual average concentration is then simply calculated by dividing the cumulative concentration for the sampled hours by the number of hours sampled arithmetic averageand the annual dry and the wet deposition fluxes are calculated by scaling the respective cumulative fluxes for the sampled hours by the ratio of the total hours to the sampled hours.
The concentrations and the dry deposition fluxes are based on the weighted contributions from the regular samples, modeled as dry hours, and the wet hour samples. The regular samples consist of all the hours based on regular sampling interval, but the effects of precipitation are ignored so that their contribution represents only dry conditions, while the contribution from the wet hour samples represents only wet conditions.
The wet deposition fluxes are only based on the wet hour samples. The following illustrates the calculation of the ANNUAL impacts when both the regular sampling as well as the wet hour sampling are selected: Optionally, the user can create an output file by specifying the Filnam parameter containing the meteorological data for the sampled hours in the same format used in the summary of the first 24 hours of data included in the main output file.
NRegStart is required to have a value from 1 through 24, i. There are no restrictions for NReglnt; however, NReglnt would generally be greater than 1.
An input of 0 zero for NWetlnt indicates that the user has not selected the wet hour sampling. This is accomplished by incorporation of a 2- point Gaussian Quadrature routine for numerical integration for some situations instead of the Romberg numerical integration utilized in the regulatory default mode.
When the AREADPLT option is specified the model will apply a single "effective" depletion factor to the undepleted area source integral, rather than applying the numerical integration for depletion within the area source integral. There are three new keywords on the CO pathway and one new keyword on the SO pathway that are used for specifying inputs for the gas dry deposition algorithm.
The inputs for these keywords are described below. The use of the gas dry deposition algorithm in ISCST3 also requires additional meteorological parameters, which can be provided by the MPRM meteorological preprocessor. The formats for the meteorological data input file for gas dry deposition applications is also described below. Specifying the State of Vegetation An optional keyword is available on the Control pathway to allow the user to specify the state of vegetation for use with the gaseous dry deposition algorithm of the ISCST3 model.
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Three options are available on this keyword, one for active and unstressed vegetation, one for active and stressed vegetation, and another for inactive vegetation.
Optional, Non-repeatable INDEX where the secondary keyword options describe the three options for the state of vegetation. The state of vegetation is used in the model, along with ambient temperature and incoming short- wave radiation, to determine the resistance to transport through the stomatal pores.
For unirrigated vegetation, the user should select the appropriate option for vegetation state based on existing soil moisture conditions.
For irrigated vegetation, the user should assume that the vegetation is active and unstressed. Option for Overriding Default Reference Parameters for Gas Dry Deposition An optional keyword is available on the Control pathway to allow the user to override the default reference parameters of cuticle resistance, ground resistance, and pollutant reactivity for use with the gas dry deposition algorithm.
Optional, Non-repeatable where the parameter Rcutr is the reference value for cuticle resistance, Rgr is the reference value for ground resistance, Reactr is the reference value for pollutant reactivity, and Refpoll is the optional name of the reference pollutant.
Option for Specifying the Deposition Velocity for Gas Dry Deposition An optional keyword is available on the Control pathway to allow the user to specify the deposition velocity for use with the gaseous dry deposition algorithm of the ISCST3 model. A single deposition velocity can be input for a given model run, and is used for all sources of gaseous pollutants.
Selection of this option will by-pass the algorithm for computing deposition velocities for gaseous pollutants, and should only be used when sufficient data to run the algorithm are not available. Results of the ISCST3 model based on a user-specified deposition velocity should be used with extra caution.
A non-fatal warning message is generated by the model if a value of Uservd greater than 0. The gas dry deposition variables may be input for a single source, or may be applied to a range of sources. The Alphas and Henry parameters are only used when applying the algorithm over a water surface. If no water surfaces are present in a particular application, then dummy non-zero values may be input for Alphas and Henry. If the deposition algorithms are being used, then the unformatted data file cannot be used.
The SEAS column is the season index, and is 1 for winter, 2 for spring, 3 for summer and 4 for fall. The records loop through hour-of- day first, and then through the seasons. Unnecessary code has been removed, and proper error handling has been implemented. The unformatted data file option has been removed due to unformatted files are not portable across different computer systems and compilers, and unformatted files cannot be used with the deposition algorithms in ISCST3.
The day-of-week variability allows for different emission factors to be specified for Weekdays Monday-FridaySaturdays, and Sundays. The user also has the option of using the Srcrng parameter for specifying a range of sources for which the emission rate factors apply, instead of identifying a single source. This is accomplished by two source ID character strings separated by a dash, e. The parameter Qflag is the variable emission rate flag, and is one of the following secondary keywords: An example of each of these options is presented below, with column headers to indicate the order in which values are to be input.
Certain individuals and offices are specifically recognized below. Valuable comments which contributed to the final report were received on the public comment draft from EPA's Science Advisory Board review and from State agencies, State organizations, and various industry groups. Additionally, valuable technical support for report development was provided by ICF Incorporated.
OAQPS staff who have contributed to the development of this document include: Mark Kataoka and Ms. Legislative Recommendations 5. Ecological Risk Assessment Framework 28 Exhibit 6: Sources of Information for Hazard Identification 47 Exhibit Cancer Dose-response Curve 57 Exhibit Examples of Chronic Toxicity Criteria 59 Exhibit Examples of Acute Toxicity Criteria 60 Exhibit Sources of Information for Ecological Effects 62 Exhibit Ozone Risk to Agroecosystems 74 Exhibit An estimate of the daily exposure that is likely to be without deleterious effect even if continued exposure occurs over a lifetime.
One dose or exposure or multiple doses or exposures occurring within a short time relative to the life of a person or other organism e. Defined in CAA section a 7 as "any significant and widespread adverse effect, which may reasonably be anticipated, to wildlife, aquatic life, or other natural resources, including adverse impacts on populations of endangered or threatened species or significant degradation of environmental quality over broad areas.
An explicit expression of the actual environmental value that is to be protected, operationally defined by an ecological entity and its attributes. For example, salmon are valued ecological entities; reproduction and age class structure are some of their important attributes.
Together "salmon reproduction and age class structure" form an assessment endpoint. An exposure level that corresponds to a predetermined level of response, such as 10 percent of test animals affected. The net accumulation of a substance by an organism as a result of uptake from all routes of exposure e. The concentration of a substance in tissue of an organism divided by its concentration in an environmental medium in situations where the organism and its food are exposed i.
The net accumulation of a substance by an organism as a result of uptake directly from an environmental medium e. The concentration of a substance in tissue of an organism divided by the concentration in an environmental medium, typically in situations where exposure is by contact or uptake directly from that medium e.
A method of statistical analysis in which the user empirically constructs sampling distributions when data are limited. Multiple exposures occurring over an extended period of time or a significant fraction of the animal's or the individual's lifetime. A condition or variable that may be a factor in producing the same response as the agent under study.
The effects of such factors may be discerned through careful design and analysis. Defined by the National Research Council as "essentially policy judgments of how to accommodate uncertainties.
They include various assumptions that are needed for assessing exposure and risk, such as scaling factors to be used for converting test responses in rodents to estimated responses in humans. The quantitative characterization of the relationship between the amount of an agent either administered, absorbed, or believed to be effective and changes in certain aspects of the biological system e.
A lifetime exposure concentration protective of adverse, non-cancer health effects that assumes all of the exposure to a contaminant is from a drinking water source. A general term that may refer to a species, a group of species, an ecosystem function or characteristic, or a specific habitat.
An ecological entity is one component of an assessment endpoint. Dose or concentration associated with a 10 percent level of response. An estimation of a numerical value of an empirical measured function at a point outside the range of data that were used to calibrate the function. The quantitative risk estimates for carcinogens are generally low dose extrapolations based on observations made at higher doses. The ratio of a level of exposure for a single substance over a specified time period to a reference level e.
Exposure concentration for humans that has been adjusted for dosimetric differences between experimental animal species and humans to be equivalent to the exposure concentration associated with observed effects in the experimental animal species.
In addition to the Met receptor family, PSI domains are also found in the plexins, Semaphorins and integrins, hence its name Bork et al. In Met, the PSI domain is connected via 4 IPT domains to the transmembrane helix and the kinase domain in the intracellular portion of the receptor. IPT domains are related to immunoglobulin-like domains and are named after their presence in plexins and transcription factors Takagi et al.
Studies involving the cross-linking of Met receptors by a variety of specific antibodies to its extracellular domain have demonstrated that simple dimerization of Met is sufficient for activation Prat et al. Based on these characteristics, the fundamental mechanism for Met dimerization remains unclear.
Coordinates of a crystal structure solved by molecular replacement are listed in Table 2. This use of the structure includes: The N-terminal amine forms a salt bridge with the side-chain of Asp and thus rearranges the loops that carry the catalytic triad.
The numbering system with a lower case c is that of the chymotyrpsinogen numbering system. Asterisks indicate amino acid residues that are conserved when the three sequences are compared and dots indicate amino acids that are conservative substitutions. Figure 2A provides a view onto the 'top' side of the propeller.
The numbers in the center refer to the blades. Disordered residues in the represented model are indicated with dotted lines and the dotted line associated numbers refer to the last and first amino acid residues present in the model. Figure 2B provides a side view of the same complex of Figure 2A. Note that the loops on the top face of the propeller are longer than the ones on the bottom face. All figures were made using Pymol DeLano, The left panel of Figure 2C captures the complex in the same view as Figure 2A and shows approximate molecular dimensions.
The right panel of Figure 2C is a view towards the bottom of the propeller and indicates a proteolysis site. The secondary structure elements depicted refer to the Met structure. The structural elements identified as A 1B 1C 1etc. The amino acids forming other blades of the propellor are also identified.
The boxes indicate structural equivalent positions between Met and Sema4D. The coordinates of the Sema3A structure were unavailable. Residues in the dimer interface of Sema4D are shaded.
Cysteines engaged in disulfide bonds are marked with letters A to G and those with same letter form a disulfide bond. Residues that are disordered in the represented structure are shown in italics.
Figure 4A shows a superposition representation of Met and Sema4D. The topology of the PSI domains in both structures is identical, but the relative orientation in comparison to the Sema domains is rather different. Figure 4B and 4C shows two different views of a model of a potential Met-dimer based on the dimer of Sema4D and the superposition shown in Figure 4 A.
The interface between the two molecules forming the Sema4D dimer is large and buries approximately 2, A2. If this dimerization interface were present on Met, the respective interface in Met would be much smaller due to the different conformation of the loops that correspond to the loops forming the dimer interface in Sema4D.
Atoms of an amino acid residues of Met shown on the right closer than about 4. Contact residues are labeled. HGF is on the left side and Met is on the right side. The three underlined amino acid residue numbers indicate the residues that form the catalytic triad in serine proteases.
Figure 5B shows differences in binding mode between a HGF: Met complex left and a trypsin: Orientations of HGF and trypsin are similar to the view in Figure 1. Contact residues of Met are shown as sticks. The numbers refer to Met residues. Glu E of Met is shown. Met complexes that may represent a portion of the active signaling complex.
Figure 8 shows a schematic diagram for Met signaling. Figure 8A shows the domain structure of Met. Figure 8C shows a 2: Figure 8D shows a 2: Such complexes may be stabilized via heparin or other co-receptors. A Angstrom AA or aa Amino acid; Amino acids are represented by single letter code or three letter code PSI domain is a small domain, which follows the Sema domain of Met, and spans about 50 residues and contains 4 disulfide bonds trypsin: The following definitions are used herein, unless specifically or contextually indicated otherwise: The term "wild type HGF sequence" generally refers to an amino acid sequence found in a naturally occurring HGF and includes naturally occurring truncated or secreted forms, variant forms e.
In some embodiments, the conversion results at least in part from cleavage between residue and residue of the wild type HGF protein. Generally, adoption of the conformation reveals a Met binding site, as described herein.
The variants include those polypeptides that have substitutions, additions or deletions. The variants also include those polypeptides that have at least one conservative amino acid substitutions, preferably all of the substitutions are conservative.
In other embodiments, the variant can bind to the Met receptor but not activate it. The term "wild-type Met receptor" generally refers to a polypeptide comprising an amino acid sequence found in a naturally occurring Met receptor and includes naturally occurring truncated or secreted forms, variant forms e.Four-Year-Old Fights Acute Lymphoblastic Leukemia
The "Met Sema domain" comprises the N terminal amino acid residues of a wild type Met receptor. A PSI domain follows the Sema domain and comprises 50 amino acid residues and has 4 disulfide bonds. IPT domains are related to immunoglobulin like domains.
The term "Met receptor variant", as used herein, refers to a polypeptide that has a different sequence than a reference polypeptide, whereine the reference polypeptide is the Met receptor that comprises an amino acid sequence of SEQ ID NO: Variants also include those polypeptides that have at least one conservative amino acid substitution, preferably, all of the substitutions are conservative. In some embodiments, the Met receptor variant has about conservative amino amino acid substitutions, more preferably about conservative amino acids substitutions, more preferably about conservative amino acid substitions, more preferably about 1 -5 conservative amino acid substitutions, and more preferably about conservative amino acid substitutions.
In some embodiments, the variant has the biological activity of binding to HGF, but not becoming activated. Thus, a binding site may include or consist of features such as cavities, surfaces, or interfaces between domains. Ligands that may associate with a binding site include, but are not limited to, cofactors, substrates, receptors, agonists, and antagonists.
A structural binding site includes "in contact" amino acid residues as determined from examination of a three-dimensional structure. Some of the "in contact" amino acid residues may not cause any change in a biochemical assay, a cell-based assay, or an in vivo assay used to define a functional binding site but may contribute to the formation of a three dimensional structure.
A functional binding site includes amino acid residues that are identified as binding site residues based upon loss or gain of function, for example, loss of binding to ligand upon mutation of the residue. In some embodiments, the amino acid residues of a functional binding site are a subset of the amino acid residues of the structural binding site.
Numbering of amino acids is that of the native receptor. A structurally equivalent "Met binding site" is defined by root mean square deviation from the structure coordinates of the amino acids that make up the binding sites in Met of at most about 0. The term " a blade of a propeller" refers to a structural feature of the Met receptor.
A blade is formed by four antiparallel strands with strand A in the center of the blade followed by strands B and C, and with strand D forming the outermost strand of the blade. The 7 blades are arranged in a circular fashion, with the N terminal strand forming strand D of the last blade.
In some embodiments, each of the blades of the propellor of the human Met receptor comprise the amino acid sequence as identified in Figure 3. The term "corresponding" or "corresponds" refers to an amino acid residue or amino acid sequence that is found at the same positions or positions in a sequence when the amino acid position or sequences are aligned with a reference sequence.
It will be appreciated that when the amino acid position or sequence is aligned with the reference sequence the numbering of the amino acids may differ from that of the reference sequence or a different numbering system may be utilized. For example, structurally homologous molecules of Met receptor include Met receptor variants, preferably variants with one or more conservative amino acid substitutions.
In some embodiments, a Met receptor variant has only conservative amino acid substitutions. Homolog tertiary structure can be probed, measured, or confirmed by known analytic or diagnostic methods, for example, X-ray, NMR, circular dichroism, a panel of monoclonal antibodies that recognize native Met receptor, and like techniques.
For example, structurally homologous molecules can have substitutions, deletions or additions of one or more contiguous or noncontiguous amino acids, such as a loop or a domain. Structurally homologous molecules also include "modified" Met receptor molecules that have been chemically or enzymatically derivatized at one or more constituent amino acid, including side chain modifications, backbone modifications, and N- and C- terminal modifications including acetylation, hydroxylation, methylation, amidation, and the attachment of carbohydrate or lipid moieties, cofactors, and like modifications.
Pharmaceutical salts can be obtained by reacting a binding-active compound of the disclosure with inorganic or organic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like.
Prodrugs are often useful because, in some situations, they may be easier to administer than the parent drug. They may, for instance, be bioavailable by oral administration whereas the parent is not.
The prodrug may also have improved solubility in pharmaceutical compositions over the parent drug. An example, without limitation, of a prodrug would be a compound of the present invention which is administered as an ester the "prodrug" to facilitate transport across a cell membrane where water solubility is detrimental to mobility but which then is metabolically hydrolyzed to the carboxylic acid, the active entity, once inside the cell where water solubility is beneficial.
A further example of a prodrug might be a short peptide polyaminoacid bonded to an acid group wherein the peptide is metabolized to yield the active moiety. A space group is designated by a capital letter identifying the lattice type P, A, F, etc. Note that the point group symmetry for a given space group can be determined by removing the cell centering symbol of the space group and replacing all screw axes by similar rotation axes and replacing all glide planes with mirror planes.
The point group symmetry for a space group describes the true symmetry of its reciprocal lattice. The quantity c is the height of the unit cell. X-ray crystallography is a technique that exploits the fact that X-rays are diffracted by crystals. X-rays have the proper wavelength in the Angstrom A range, approximately 10"8 cm to be scattered by the electron cloud of an atom of comparable size.
Based on the diffraction pattern obtained from X-ray scattering of the periodic assembly of molecules or atoms in the crystal, the electron density can be reconstructed. Additional phase information can be extracted either from the diffraction data or from supplementing diffraction experiments to complete the reconstruction the phase problem in crystallography. A model is then progressively built into the experimental electron density, refined against the data to produce an accurate molecular structure.
X-ray structure coordinates define a unique configuration of points in space. A similar or identical configuration can be defined by an entirely different set of coordinates, provided the distances and angles between coordinates remain essentially the same.
In addition, a configuration of points can be defined by increasing or decreasing the distances between coordinates by a scalar factor, while keeping the angles essentially the same. Modes for Carrying Out the Invention: The present disclosure also includes the three-dimensional configuration of points derived from the structure coordinates of at least a portion of an extracellular fragment of a Met receptor molecule or molecular complex, as well as structurally equivalent configurations, as described below.
Met complex binding site. The disclosure also includes the three-dimensional configuration of points identifying other structural features of an extracellular fragment of the Met receptor. Those other structural features include the blades of the propeller structure and PSI domain.
A plurality of amino acid residues have been identified as contributing to these structural features of Met receptor. In some embodiments, the amino acid residues comprise those identified as corresponding to structural features as shown in Figure 3. Met complex or extracellular fragment of the Met receptor including the Sema domain, as well as structurally equivalent configurations.
Structurally homologous molecules or molecular complexes are defined below. Met complex or extracellular fragment of the Met receptor according to a method of the disclosure.
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The configurations of points in space derived from structure coordinates according to the disclosure can be visualized as, for example, a holographic image, a stereodiagram, a model, or a computer-displayed image, and the disclosure thus includes such images, diagrams or models. Hepatocyte growth factor comprises a 69kDa alpha chain and 34kDa beta chain.
The 69 kDa alpha chain comprise a N terminal finger domain and four kringle domains K1-K4. The sequence of Table 4 has one amino acid change from wild type shown in Table 8; the cysteine at amino acid position is changed to a serine. Numbers in brackets or preceeded by a lower case c represent a numbering system based on reference to chymotrypsinogen. The Met receptor is a tyrosine kinase and is part of a larger family of growth factor receptors with domain architecture similar to the Ron and Sea receptors.
The extracellular portion of the Met receptor comprises N-terminal amino acids that fold into a Sema domain. A PSI domain follows the Sema domain and comprises about 50 amino acids and has 4 disulfide bonds. Crosslinking of Met receptors to form dimers also activates the Met receptor. The numbering system of Met receptor is that of the Swiss Prot database as shown in Table 6. Native or wild- type polypeptides include naturally occurring variants, secreted or truncated forms.
Several isoforms of HGF are known such as isoform 1, isoform 2, isoform 3, isoform 4, and isoform 5. The present disclosure also includes a polypeptide comprising, consisting essentially of, or consisting of a portion or fragment of the Met receptor. The polypeptide fragment includes amino acid residues from any of amino acid 1 to 25 residues to amino acid position or residues corresponding to those positions.
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In some embodiments, the polypeptide portion has the ability to bind to ligand HGF. The polypeptide portion may also be fused to heterologous polypeptide, such as a peptide tag. Preferably, the fusion polypeptide retains the ability to bind a ligand, such as HGF.
The disclosure also provides a polypeptide comprising, consisting essentially of, or consisting of a portion or fragment of the Met receptor starting at amino acid residue and ending at any one of amino acid residues to or residues corresponding to these residues.
The polypeptide portion or fragment may be fused to a heterologous polypeptide. The present disclosure also includes variants of the Met receptor. Variants include those polypeptides that have amino acid substitutions, deletions, and additions.
Amino acid substitutions can be made for example to replace cysteines and eliminate formation of disulfide bonds. Amino acid substitutions can also be made to change proteolytic cleavage sites.
The variants also include those polypeptides that have at least one conservative amino acid substitution. In some embodiments a variant only has conservative amino acid substitutions. In some embodiments, the Met receptor variant has about 1 conservative amino acid substitutions, more preferably about 1 conservative amino acids substitutions, more preferably about conservative amino acid substitutions, more preferably about 1 -5 conservative amino acid substitutions, and more preferably about 1 -2 conservative amino acid substitutions.
Fusion Proteins A Met receptor, variant or structural homolog or portions thereof, may be fused to a heterologous polypeptide or compound. The heterologous polypeptide is a polypeptide that has a different function than that of the Met receptor. Examples of heterologous polypeptide include polypeptides that may act as carriers, may extend half life, may act as epitope tags, may provide ways to detect or purify the fusion protein.
Heterologous polypeptides include KLH, albumin, salvage receptor binding epitopes, immunoglobulin constant regions, and peptide tags. Peptide tags useful for detection or purification include FLAG, gD protein, polyhistidine tags, hemagluthinin from influenza virus, T7 tag, S tag, Strep tag, chloramiphenicol acetyl transferase, biotin, glutathione-S transferase, green fluorescent protein and maltose binding protein.