Structural characterization of the latent complex between transforming growth factor β1 and β1-latency-associated peptide
| Autor: | Larry E. Gentry, Grainne A. McMAHON, John David Dignam |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
DNA
Complementary Glycosylation medicine.medical_treatment Carbohydrates Peptide CHO Cells Biochemistry Protein Structure Secondary Cell Line Transforming Growth Factor beta1 chemistry.chemical_compound Transforming Growth Factor beta Cricetinae medicine Animals Chemical Precipitation Protein Precursors Beta (finance) Lung Molecular Biology chemistry.chemical_classification Mannosephosphates biology Molecular mass Circular Dichroism Chinese hamster ovary cell Growth factor digestive oral and skin physiology Gene Amplification Proteins Cell Biology Transforming growth factor beta Chromatography Ion Exchange Molecular biology Peptide Fragments Tetrahydrofolate Dehydrogenase chemistry Mink Cell culture biology.protein human activities Cell Division Research Article |
| Zdroj: | Biochemical Journal. 313:343-351 |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/bj3130343 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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