Tyrosine-rich acidic matrix protein (TRAMP) accelerates collagen fibril formation in vitro

• Published in: The Journal of biological chemistry 1993-09, Vol.268 (26), p.19826-19832
• Main Authors: MACBEATH, J. R. E, SHACKLETON, D. R, HULMES, D. J. S
• Format: Article
• Language: English
• Subjects: Animals; Biochemistry, Molecular Biology; Biological and medical sciences; Cell coat. Cell surface; Cell structures and functions; Chondroitin Sulfate Proteoglycans - isolation & purification; Chondroitin Sulfate Proteoglycans - metabolism; Collagen - isolation & purification; Collagen - metabolism; Collagen - ultrastructure; Electrophoresis, Disc; Extracellular Matrix Proteins - isolation & purification; Extracellular Matrix Proteins - metabolism; Fundamental and applied biological sciences. Psychology; Kinetics; Lathyrism - metabolism; Life Sciences; Male; Microscopy, Electron; Molecular and cellular biology; Nephelometry and Turbidimetry; Protein-Lysine 6-Oxidase - isolation & purification; Rats; Rats, Sprague-Dawley; Skin - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/s0021-9258(19)36588-3

Tyrosine-rich acidic matrix protein (TRAMP) is a recently discovered protein that co-purifies with porcine skin lysyl oxidase and is equivalent to the M(r) 22,000 extracellular matrix protein from bovine skin that co-purifies with dermatan sulfate proteoglycans (Cronshaw, A. D., MacBeath, J. R. E., Shackleton, D. R., Collins, J. F., Fothergill-Gilmore, L. A., and Hulmes, D. J. S. (1993) Matrix 13, 255-266; Neame, P. J., Choi, H. U., and Rosenberg, L. C. (1989) J. Biol. Chem. 264, 5474-5479). The effect of TRAMP on collagen fibril formation was studied in vitro by reconstitution of fibrils from lathyritic rat skin collagen I. Fibril formation was initiated by the warm start procedure, in which acidic collagen solutions and double strength neutral buffer, both preincubated separately at 34 degrees C, were mixed. When monitored by turbidimetry, TRAMP was found to accelerate collagen fibril formation. Acceleration occurred at sub-stoichiometric molar ratios of TRAMP collagen, and the presence of TRAMP stabilized the fibrils against low temperature dissociation. It was confirmed by centrifugation that the amount of fibrillar collagen formed in the presence of TRAMP was greater than in its absence. By SDS-polyacrylamide gel electrophoresis and scanning densitometry, binding of TRAMP to collagen was detected that approached saturation with a molar ratio of TRAMP to collagen of approximately 1:2. Fibrils formed in the presence of TRAMP were normal when observed by electron microscopy, although fibril diameters were smaller than the controls. TRAMP was found to partially reverse the inhibitory effects of urea and increased ionic strength on the kinetics of fibril formation, although inhibition by glucose was unaffected. TRAMP also accelerated the assembly of pepsin-treated collagen, where the non-helical, telopeptide regions were partially removed. Acceleration of collagen fibril formation by TRAMP is discussed in the light of the known effects of other extracellular matrix components on this process.