Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Blood proteins or globulins – e.g. – proteoglycans – platelet...
Reexamination Certificate
1995-05-23
2001-03-13
Huff, Sheela (Department: 1642)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Blood proteins or globulins, e.g., proteoglycans, platelet...
C530S387900, C530S389200, C435S331000, C435S334000
Reexamination Certificate
active
06201108
ABSTRACT:
DESCRIPTION
Background
Transforming growth factor-beta (TGF-&bgr;) is a member of a family of structurally related cytokines that elicit a variety of responses, including growth, differentiation, and morphogenesis, in many different cell types. (Roberts, A. B. and M. B. Sporn, In:
Peptide Growth Factors and Their Receptors
, Springer-Verlag, Heidelberg, pp. 421-472 (1990); Massague, J.,
Annu. Rev. Cell. Biol
. 6:597-641 (1990)) In vertebrates at least five different forms of TGF-&bgr;, termed TGF-&bgr;1 to TGF-&bgr;5, have been identified; they all share a high degree (60%-80%) of amino-acid sequence identity. While TGF-&bgr;1 was initially characterized by its ability to induce anchorage-independent growth of normal rat kidney cells, its effects on most cell types are anti-mitogenic. (Altschul, S. F. et al.,
J. Mol. Biol
. 215:403-410 (1990); Andres, J. L. et al.,
J. Cell. Biol
. 109:3137-3145 (1989)) It is strongly growth-inhibitory for many types of cells, including both normal and transformed epithelial, endothelial, fibroblast, neuronal, lymphoid, and hematopoietic cells. In addition, TGF-&bgr; plays a central role in regulating the formation of extracellular matrix and cell-matrix adhesion processes.
In spite of its widespread effects on cell phenotype and physiology, little is known about the biochemical mechanisms that enable TGF-&bgr; family members to elicit these varied responses. Three distinct high-affinity cell-surface TGF-&bgr;-binding proteins, termed type I, II and III, have been identified by incubating cells with radiolabelled TGF-&bgr;1, cross-linking bound TGF-&bgr;1 to cell surface molecules, and analyzing the labelled complexes by polyacrylamide gel electrophoresis. (Massague, J. and B. Like,
J. Biol. Chem
. 260:2636-2645 (1985); Cheifetz, S. et al.
J. Biol. Chem
. 261:9972-9978 (1986).) The binding constants are about 5-50 pM for the type I and II receptor and 30-300 pM for the type III receptor. (Boyd, F. T. and J. Massague,
J. Biol. Chem
. 264:2272-2278 (1989))
The type I and II receptors, of estimated 53 and 70-100 kilodaltons mass respectively, are N-glycosylated transmembrane proteins that are similar in many respects. Each of these receptors has a distinct affinity for each member of the TGF-&bgr; family of ligands. (Boyd, F. T. and J. Massague,
J. Biol. Chem
. 264:2272-2278 (1989)) In contrast, the type III receptor shows comparable affinities for all TGF-&bgr; isotypes; the type III receptor is the most abundant cell-surface receptor for TGF-&bgr; in many cell lines (upwards of 200,000 per cell), and is an integral membrane proteoglycan. It is heavily modified by glycosaminoglycan (GAG) groups, and migrates heterogeneously upon gel electrophoresis as proteins of 280 to 330 kilodaltons. When deglycosylated with heparitinase and chondrontinase, the protein core migrates as a 100-110 kilodalton protein. The TGF-&bgr; binding site resides in this protein core, as non-glycosylated forms of this receptor that are produced in cell mutants defective in GAG synthesis are capable of ligand binding with affinities comparable to those of the natural receptor. (Cheifetz, S. and J. Massague,
J. Biol. Chem
., 264:12025-12028 (1989) A variant form of type III receptor is secreted by some types of cells as a soluble molecule that apparently lacks a membrane anchor. This soluble species is found in low amounts in serum and in extracellular matrix.
The type III receptor, also called betaglycan, has a biological function distinct from that of the type I and II receptors. Some mutant mink lung epithelial cell (Mv1Lu) selected for loss of TGF-&bgr; responsiveness no longer express type I receptors; others, similarly selected, lose expression of both the type I and II receptors. However, all these variants continue to express the type III receptor. (Boyd, F. T. and J. Massague,
J. Biol. Chem
. 264:2272-2278 (1989); Laiho, M. et al.,
J. Biol. Chem
. 265:18518-18524 (1990)) This has led to the proposal that types I and II receptors are signal-transducing molecules while the type III receptor, may subserve some other function, such as in concentrating ligand before presentation to the bona fide signal-transducing receptors. The secreted form of type III receptor, on the other hand, may act as a reservoir or clearance system for bioactive TGF-&bgr;.
Additional information about each of these TGF-&bgr; receptor types would enhance our understanding of their roles and make it possible, if desired, to alter their functions.
SUMMARY OF THE INVENTION
The present invention relates to isolation, sequencing and characterization of DNA encoding the TGF-&bgr; type III receptor of mammalian origin and DNA encoding the TGF-&bgr; type II receptor of mammalian origin. It also relates to the encoded TGF-&bgr; type III and type II receptors, as well as to the soluble form of each; uses of the receptor-encoding genes and of the receptors themselves; antibodies specific for TGF-&bgr; type III receptor and antibodies specific for TGF-&bgr; type II receptor. In particular, it relates to DNA encoding the TGF-&bgr; type III receptor of rat and human origin, DNA encoding the TGF-&bgr; type II receptor of human origin and homologues of each.
The TGF-&bgr; receptor-encoding DNA of the present invention can be used-to identify equivalent TGF-&bgr; receptor type III and type II genes from other sources, using, for example, known hybridization-based methods or the polymerase chain reaction. The type III receptor gene, the type II receptor gene or their respective encoded products can be used to alter the effects of TGF-&bgr; (e.g., by altering receptivity of cells to TGF-&bgr; or interfering with binding of TGF-&bgr; to its receptor), such as its effects on cell proliferation or growth, cell adhesion and cell phenotype. For example, the TGF-&bgr; receptor type III gene, the TGF-&bgr; receptor type II gene, or a truncated gene which encodes less than the entire receptor (e.g., soluble TGF-&bgr; type III receptor, soluble TGF-&bgr; type II receptor or the TGF-&bgr; type III or type II binding site) can be administered to an individual in whom TGF-&bgr; effects are to be altered. Alternatively, the TGF-&bgr; type III receptor, the TGF-&bgr; type II receptor, a soluble form thereof (i.e., a form lacking the membrane anchor) or an active binding site of the TGF-&bgr; type III or the type II receptor can be administered to an individual to alter the effects of TGF-&bgr;.
Because of the many roles TGF-&bgr; has in the body, availability of the TGF-&bgr; receptors described herein makes it possible to further assess TGF-&bgr; function and to alter (enhance or diminish) its effects.
REFERENCES:
patent: 5340925 (1994-08-01), Lioubin et al.
patent: 0 369 861 A1 (1990-05-01), None
Andres, J.L. et al. 1989. J. Cell Biology, 109:3137-3145.
Boyd, F.T. et al. 1990. J. Cell Sci. Suppl. 13:131-138.
Massagué, J. 1990. Annu. Rev. Cell Biol. 6: 597-641.
Goding, J.W. (ed) 1986. Monoclonal antibodies: Principles-Practice.
Campbell, Monoclonal Antibody Technology. Elsevier Science Pub. Chapter 1, 1984.
Massague, et al., “Cellular Receptors for Type Beta Transforming Growth Factor”,J. Biol. Chem.260(5):2636-2645 (1985).
Massague, J., “Subunit Structure of a High-affinity receptor for Type Beta-transforming Growth Factor”,J. Biol. Chem.,260(11):7059-7066 (1985).
Cheifetz, S., et al., “Cellular Distribution of Type I and Type II Receptors for Transforming Growth Factor-Beta*”,J. Biol. Chem.,261(21):9972-9978 (1986).
Fanger, B.O., et al., “Structure and Properties of the Cellular Receptor for Transforming Growth Factor Type Beta”,Biochem.,25:3083-3091 (1986).
Cheifetz, S., et al., “The Transforming Growth Factor-Beta System, a Complex Pattern of Cross-Reactive Ligands and Receptors”,Cell,48:409-415 (1987).
Ignotz, R.A. et al., “Cell Adhesion Protein Receptors as Targets for Transforming Growth Factor-Beta Action”,Cell,51:189-197 (1987).
Segarini, P.R., et al., “The High Molecular Weight Receptor to Transforming Growth Factor-Beta Contains Glycosaminoglycan Chains”,J. Biol. Chem.,263(17):8366-8370 (1988).
Chiefetz
Lin Herbert Y.
Lodish Harvey F.
Wang Xiao-Fan
Weinberg Robert A.
Hamilton Brook Smith & Reynolds P.C.
Huff Sheela
Whitehead Institute for Medical Research
LandOfFree
TGF-&bgr; type receptor cDNAs encoded products and uses... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with TGF-&bgr; type receptor cDNAs encoded products and uses..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and TGF-&bgr; type receptor cDNAs encoded products and uses... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2520557