Release of biological activities from quiescent fibronectin by a conformational change and limited proteolysis by matrix metalloproteinases.
We reported that specific biological activities are confined to three domains of the fibronectin (Fn) molecule [Fukai et al. (1991) J. Biol. Chem. 266, 8807; Fukai et al. (1993) Biochemistry 32, 5746]: the potent ability to stimulate the adipocyte differentiation of ST-13 cells is in the amino-terminal fibrin-binding (Fib 1) domain (referred to as Fib 1 domain activity); the RGD-dependent activities that stimulate NIH-L13 cell migration and inhibit adipocyte differentiation are in the central cell-binding (Cell) domain (Cell domain activity); and the activity that stimulates cell migration in a RGD-independent manner is in the carboxyl-terminal fibrin-binding (Fib 2) domain (Fib 2 domain activity). Human plasma Fn which was purified without exposure to a denaturant, such as urea, exhibited no Fib 1, Fib 2, or Cell domain activity. By exposure to urea or surface adsorption, Fn showed Cell domain activity but not those of the Fib 1 and Fib 2 domains. Whether the cryptic domain activities are disclosed or not depended on whether or not the responsible domains were irreversibly exposed from confined environments of Fn structure as confirmed by light-scattering measurement and enzyme immunoassay using domain-specific monoclonal antibodies. We then investigated the action of matrix metalloproteinases (MMPs) in liberating the Fib 1, Cell, and Fib 2 domain activities. Matrilysin released only the Cell domain activity. In contrast, stromelysin, collagenase, and especially gelatinase A additionally liberated the Fib 1 and Fib 2 domain activities. The Fib 1, Fib 2, and Cell domains acquired much higher activities when they were freed from linkage with adjacent domains.(ABSTRACT TRUNCATED AT 250 WORDS)
Fukai
F
F
Department of Patho-Physiology, Faculty of Pharmaceutical Sciences, Science University of Tokyo.
Ohtaki
M
M
Fujii
N
N
Yajima
H
H
Ishii
T
T
Nishizawa
Y
Y
Miyazaki
K
K
Katayama
T
T
D000595:Amino Acid Sequence; D002460:Cell Line; D005109:Extracellular Matrix / Q000201:enzymology; D005353:Fibronectins / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis; D008666:Metalloendopeptidases / Q000378:metabolism*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism; D011487:Protein Conformation
D005353:Fibronectins; D009842:Oligopeptides; C047981:arginyl-glycyl-aspartic acid; D008666:Metalloendopeptidases