Involvement of fibronectin type II repeats in the efficient inhibition of gelatinases A and B by long-chain unsaturated fatty acids.
The matrix metalloproteinases gelatinase A (MMP-2) and gelatinase B (MMP-9) are implicated in the physiological and pathological breakdown of several extracellular matrix proteins. In the present study, we show that long-chain fatty acids (e.g. oleic acid, elaidic acid, and cis- and trans-parinaric acids) inhibit gelatinase A as well as gelatinase B with K(i) values in the micromolar range but had only weak inhibitory effect on collagenase-1 (MMP-1), as assessed using synthetic or natural substrates. The inhibition of gelatinases depended on fatty acid chain length (with C18 > C16, C14, and C10), and the presence of unsaturations increased their inhibitory capacity on both types of gelatinase. Ex vivo experiments on human skin tissue sections have shown that micromolar concentrations of a long-chain unsaturated fatty acid (elaidic acid) protect collagen and elastin fibers against degradation by gelatinases A and B, respectively. In order to understand why gelatinases are more susceptible than collagenase-1 to inhibition by long-chain fatty acids, the possible role of the fibronectin-like domain (a domain unique to gelatinases) in binding inhibitory fatty acids was investigated. Affinity and kinetic studies with a recombinant fibronectin-like domain of gelatinase A and with a recombinant mutant of gelatinase A from which this domain had been deleted pointed to an interaction of long-chain fatty acids with the fibronectin-like domain of the protease. Surface plasmon resonance studies on the interaction of long-chain fatty acids with the three individual type II modules of the fibronectin-like domain of gelatinase A revealed that the first type II module is primarily responsible for binding these compounds.
CNRS FRE 2260, IFR 53 Biomolecules, Faculty of Medicine, 51 Rue Cognacq Jay, F-51100 Reims, France.
A
A
Berton
V
V
Rigot
E
E
Huet
M
M
Decarme
Y
Y
Eeckhout
L
L
Patthy
G
G
Godeau
W
W
Hornebeck
G
G
Bellon
H
H
Emonard
D003094:Collagen / Q000378:metabolism; D004549:Elastin / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000737:chemistry / Q000494:pharmacology*; D005353:Fibronectins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D061965:Matrix Metalloproteinase Inhibitors; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid; D012867:Skin / Q000201:enzymology / Q000378:metabolism
D005231:Fatty Acids, Unsaturated; D005353:Fibronectins; D061965:Matrix Metalloproteinase Inhibitors; D011994:Recombinant Proteins; D003094:Collagen; D004549:Elastin