Structural Characterization of the Latent Complex Between Transforming Growth Factor Beta 1 and Beta 1-latency-associated Peptide

Overview

Journal Biochem J

Specialty Biochemistry

Date 1996 Jan 1

PMID 8546705

Citations 26

Authors

G A McMahon

J D Dignam

L E Gentry

Affiliations

Soon will be listed here.

Abstract

The formation of a non-covalent complex between mature transforming growth factor beta 1 (TGF-beta 1) and its pro region, the beta 1-latency-associated peptide (beta 1-LAP), is important in regulating the activity of this multipotent growth factor. We have overexpressed simian beta 1-LAP in Chinese hamster ovary (CHO) cells to produce a cell line which secretes beta 1-LAP into the culture medium at > 1 mg/l, thus enabling structural studies of complex formation between beta 1-LAP and TGF-beta 1. The simian beta 1-LAP expressed in CHO cells reversed the growth inhibitory effect of exogenous TGF-beta 1 on Mv1Lu (mink lung epithelial) cells and was able to form a cross-linked complex with 125I-TGF-beta 1. Simian beta 1-LAP was purified to homogeneity by a combination of ammonium sulphate precipitation, gel filtration, dye ligand chromatography and anion-exchange chromatography, with a yield of 15%. The purified protein had an apparent molecular mass of 114 kDa as determined by SDS/PAGE, which is greater than that determined for the transient expression of simian beta 1-LAP in COS-1 and for the simian precursor of TGF-beta 1 (pro-TGF-beta 1) in CHO cells, this major difference being due to more extensive glycosylation of beta 1-LAP expressed by this CHO clone. Far-UV CD spectroscopy of simian beta 1-LAP indicates a mostly beta-sheet structure, with extensive structural rearrangements occurring upon formation of the latent complex between TGF-beta 1 and beta 1-LAP.

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References

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Daopin S, Piez K, Ogawa Y, Davies D . Crystal structure of transforming growth factor-beta 2: an unusual fold for the superfamily. Science. 1992; 257(5068):369-73. DOI: 10.1126/science.1631557. View

De Larco J, Todaro G . Growth factors from murine sarcoma virus-transformed cells. Proc Natl Acad Sci U S A. 1978; 75(8):4001-5. PMC: 392918. DOI: 10.1073/pnas.75.8.4001. View

Smith D, Fisher P . Identification, developmental regulation, and response to heat shock of two antigenically related forms of a major nuclear envelope protein in Drosophila embryos: application of an improved method for affinity purification of antibodies using.... J Cell Biol. 1984; 99(1 Pt 1):20-8. PMC: 2275611. DOI: 10.1083/jcb.99.1.20. View

Kaufman R, Wasley L, Spiliotes A, Gossels S, LATT S, Larsen G . Coamplification and coexpression of human tissue-type plasminogen activator and murine dihydrofolate reductase sequences in Chinese hamster ovary cells. Mol Cell Biol. 1985; 5(7):1750-9. PMC: 367294. DOI: 10.1128/mcb.5.7.1750-1759.1985. View

Derynck R, Jarrett J, Chen E, EATON D, Bell J, Assoian R . Human transforming growth factor-beta complementary DNA sequence and expression in normal and transformed cells. Nature. 1985; 316(6030):701-5. DOI: 10.1038/316701a0. View

Lawrence D, Pircher R, Jullien P . Conversion of a high molecular weight latent beta-TGF from chicken embryo fibroblasts into a low molecular weight active beta-TGF under acidic conditions. Biochem Biophys Res Commun. 1985; 133(3):1026-34. DOI: 10.1016/0006-291x(85)91239-2. View

Ruoslahti E, Pierschbacher M . Arg-Gly-Asp: a versatile cell recognition signal. Cell. 1986; 44(4):517-8. DOI: 10.1016/0092-8674(86)90259-x. View

Yang J, Wu C, Martinez H . Calculation of protein conformation from circular dichroism. Methods Enzymol. 1986; 130:208-69. DOI: 10.1016/0076-6879(86)30013-2. View

10.

Sharples K, Plowman G, Rose T, Twardzik D, Purchio A . Cloning and sequence analysis of simian transforming growth factor-beta cDNA. DNA. 1987; 6(3):239-44. DOI: 10.1089/dna.1987.6.239. View

11.

Ikeda T, Lioubin M, Marquardt H . Human transforming growth factor type beta 2: production by a prostatic adenocarcinoma cell line, purification, and initial characterization. Biochemistry. 1987; 26(9):2406-10. DOI: 10.1021/bi00383a002. View

12.

Gentry L, Webb N, Lim G, Brunner A, Ranchalis J, Twardzik D . Type 1 transforming growth factor beta: amplified expression and secretion of mature and precursor polypeptides in Chinese hamster ovary cells. Mol Cell Biol. 1987; 7(10):3418-27. PMC: 367992. DOI: 10.1128/mcb.7.10.3418-3427.1987. View

13.

Miyazono K, Hellman U, Wernstedt C, Heldin C . Latent high molecular weight complex of transforming growth factor beta 1. Purification from human platelets and structural characterization. J Biol Chem. 1988; 263(13):6407-15. View

14.

Wakefield L, Smith D, Flanders K, Sporn M . Latent transforming growth factor-beta from human platelets. A high molecular weight complex containing precursor sequences. J Biol Chem. 1988; 263(16):7646-54. View

15.

Lyons R, Keski-Oja J, Moses H . Proteolytic activation of latent transforming growth factor-beta from fibroblast-conditioned medium. J Cell Biol. 1988; 106(5):1659-65. PMC: 2115066. DOI: 10.1083/jcb.106.5.1659. View

16.

Brunner A, Gentry L, Cooper J, Purchio A . Recombinant type 1 transforming growth factor beta precursor produced in Chinese hamster ovary cells is glycosylated and phosphorylated. Mol Cell Biol. 1988; 8(5):2229-32. PMC: 363407. DOI: 10.1128/mcb.8.5.2229-2232.1988. View

17.

Purchio A, Cooper J, Brunner A, Lioubin M, Gentry L, Kovacina K . Identification of mannose 6-phosphate in two asparagine-linked sugar chains of recombinant transforming growth factor-beta 1 precursor. J Biol Chem. 1988; 263(28):14211-5. View

18.

Gentry L, Lioubin M, Purchio A, Marquardt H . Molecular events in the processing of recombinant type 1 pre-pro-transforming growth factor beta to the mature polypeptide. Mol Cell Biol. 1988; 8(10):4162-8. PMC: 365485. DOI: 10.1128/mcb.8.10.4162-4168.1988. View

19.

Miyazono K, Heldin C . Role for carbohydrate structures in TGF-beta 1 latency. Nature. 1989; 338(6211):158-60. DOI: 10.1038/338158a0. View

20.

Kovacina K, Purchio A, Lioubin M, Miyazono K, Heldin C, Roth R . Interactions of recombinant and platelet transforming growth factor-beta 1 precursor with the insulin-like growth factor II/mannose 6-phosphate receptor. Biochem Biophys Res Commun. 1989; 160(1):393-403. DOI: 10.1016/0006-291x(89)91669-0. View