US5340925A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Normal human growth regulatory receptor for TGF-β
Assignee: Oncogen LP
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Filing: 1992-06-17
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Granted: 1994-08-23
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Expires: 2012-06-17
Abstract
Abstract Type III TGF-β receptor is identified in and purified from normal human embryonic palatal mesenchyme (HEPM) cells and the purified product characterized structurally and functionally. HEPM cells were found to express high levels of the type III TGF-β receptor and were found to significantly down-regulate two classes of TGF-β receptor binding site. Purification of the type III TGF-β receptor from solubilized HEPM cell membranes by affinity chromatography yielded a biologically active protein of about 205 kd which specifically binds both the recombinant and natural forms of TGF-β1 and TGF-β2, with affinity dissociation constants in the picomolar range.
Inventors
- 👤 Mario N. Lioubin
- 👤 Thomas J. Brown
- 👤 Anthony F. Purchio
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 07:50
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/705; C07K14/71
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
85 words
A protein that: (a) has an amino-terminal amino acid sequence comprising: NH.sub.2 -Lys-Tyr-Tyr-Asp-Lys-Asp-Tyr; (b) has a glycosylated apparent molecular weight by gel electrophoresis under reducing conditions of about 200,000 to about 206,000 daltons; (c) may be obtained from a naturally occurring mammalian fibroblast; (d) is a specific receptor for TGF-β; and (e) is free from bands due to other proteins which occur naturally in mammalian fibroblasts, when said protein is analyzed by the polyacrylamide gel electrophoresis procedure of Laemmli using Coomassie stain for protein detection.
Claim 2
Dependent Claim
85 words
A protein that: (a) has an amino-terminal amino acid sequence comprising: NH.sub.2 -Lys-Tyr-Tyr-Asp-Lys-Asp-Tyr; (b) has a glycosylated apparent molecular weight by gel electrophoresis under reducing conditions of about 200,000 to about 206,000 daltons; (c) may be obtained from a naturally occurring mammalian fibroblast; (d) is a specific receptor for TGF-β; and (e) is free from bands due to other proteins which occur naturally in mammalian fibroblasts, when said protein is analyzed by the polyacrylamide gel electrophoresis procedure of Laemmli using Coomassie stain for protein detection.
Claim 3
Dependent Claim
6 words
A glycosylated protein of claim 1.
Claim 4
Dependent Claim
6 words
An un-glycosylated protein of claim 1.
Claim 5
Dependent Claim
11 words
The protein of claim 2 in which the TGF-β is TGF-β1.
Claim 6
Dependent Claim
12 words
The protein of claim 4 in which the TGF-β1 is recombinant TGF-β1.
Claim 7
Dependent Claim
11 words
The protein of claim 2 in which the TGF-β is TGF-β2.
Claim 8
Dependent Claim
12 words
The protein of claim 2 in which the TGF-β is recombinant TGF-β2.
Claim 9
Dependent Claim
21 words
The protein of claim 5 having a dissociation constant for TGF-β1 of about 1×10 -12 M to about 1×10 -9 M.
Claim 10
Dependent Claim
149 words
A Type III TGF-β receptor, obtained by the method comprising: (a) solubilizing membranes from cultures HEPM cells to make a solubilized HEPM membrane preparation; (b) covalently binding TGF-β to a solution-permeable matrix to make a TGF-β affinity matrix; (c) contacting the solubilized HEPM membrane preparation with the TGF-β affinity matrix; (d) removing unbound components of the solubilized HEPM membrane preparation from the TGF-β affinity matrix; and (e) eluting said Type III TGF-β receptor from said TGF-β affinity matrix; (f) has an amino-terminal amino acid sequence comprising NH.sub.2 -Lys-Tyr-Tyr-Asp-Lys-Asp-Tyr; (g) has a glycosylated apparent molecular weight by gel electrophoresis under reducing conditions of about 200,000 to 260,000 daltons; (h) specifically binds TGF-β; and (i) is free from bands due to other proteins which occur naturally in mammalian fibroblasts, when said Type III TGF-β receptor is analyzed by the polyacrylamide gel electrophoresis procedure of Laemmli using Coomassie stain for protein detection.
Claim 11
Dependent Claim
149 words
A Type III TGF-β receptor, obtained by the method comprising: (a) solubilizing membranes from cultures HEPM cells to make a solubilized HEPM membrane preparation; (b) covalently binding TGF-β to a solution-permeable matrix to make a TGF-β affinity matrix; (c) contacting the solubilized HEPM membrane preparation with the TGF-β affinity matrix; (d) removing unbound components of the solubilized HEPM membrane preparation from the TGF-β affinity matrix; and (e) eluting said Type III TGF-β receptor from said TGF-β affinity matrix; (f) has an amino-terminal amino acid sequence comprising NH.sub.2 -Lys-Tyr-Tyr-Asp-Lys-Asp-Tyr; (g) has a glycosylated apparent molecular weight by gel electrophoresis under reducing conditions of about 200,000 to 260,000 daltons; (h) specifically binds TGF-β; and (i) is free from bands due to other proteins which occur naturally in mammalian fibroblasts, when said Type III TGF-β receptor is analyzed by the polyacrylamide gel electrophoresis procedure of Laemmli using Coomassie stain for protein detection.
Claim 12
Dependent Claim
11 words
The protein of claim 9 in which the TGF-β is TGF-β1.
Claim 13
Dependent Claim
12 words
The protein of claim 10 in which the TGF-β is recombinant TGF-β1.
Claim 14
Dependent Claim
11 words
The protein of claim 9 in which the TGF-β is TGF-β2.
Claim 15
Dependent Claim
12 words
The protein of claim 12 in which the TGF-β is recombinant TGF-β2.
Claim 16
Dependent Claim
21 words
The protein of claim 11 having a dissociation constant for TGF-β1 of about 1×10 -12 M to about 1×10 -9 M.
Claim 17
Dependent Claim
12 words
The protein of claim 1 wherein said fibroblast is an HEPM cell.
References Cited (Backward Citations)
US4286690A
Cited
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