Apparent tradeoff of higher activity in MMP-12 for enhanced stability and flexibility in MMP-3.
The greater activity of MMP-12 than MMP-3 toward substrates from protein fibrils has been quantified. Why is MMP-12 the more active protease? We looked for behaviors associated with the higher activity of MMP-12 than MMP-3, using nuclear magnetic resonance to monitor backbone dynamics and residue-specific stabilities of their catalytic domain. The proteolytic activities are likely to play important roles in inflammatory diseases of arteries, lungs, joints, and intestines. Nuclear magnetic resonance line broadening indicates that regions surrounding the active sites of both proteases sample conformational substates within milliseconds. The more extensive line broadening in MMP-3 suggests greater sampling of conformational substates, affecting the full length of helix B and beta-strand IV forming the active site, and more remote sites. This could suggest more excursions to functionally incompetent substates. MMP-3 also has enhanced subnanosecond fluctuations in helix A, in the beta-hairpin of strands IV and V, and before and including helix C. Hydrogen exchange protection in the EX2 regime suggests that MMP-3 possesses 2.8 kcal/mol higher folding stability than MMP-12(E219A). The beta-sheet of MMP-3 appears to be stabilized still more. The higher stability of MMP-3 relative to MMP-12 coincides with the former's considerably lower proteolytic activity. This relationship is consistent with the hypothesis that enzymes often trade stability for higher activity.
Liang
Xiangyang
X
Department of Biochemistry, University of Missouri, Columbia, Missouri, USA.
Arunima
A
A
Zhao
Yingchu
Y
Bhaskaran
Rajagopalan
R
Shende
Anuradha
A
Byrne
Todd S
TS
Fleeks
Jeremy
J
Palmier
Mark O
MO
Van Doren
Steven R
SR
D020134:Catalytic Domain; D041961:Deuterium Exchange Measurement; D004795:Enzyme Stability; D006801:Humans; D007700:Kinetics; D053508:Matrix Metalloproteinase 12 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019278:Matrix Metalloproteinase 3 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D009068:Movement; D009154:Mutation
D019278:Matrix Metalloproteinase 3; D053508:Matrix Metalloproteinase 12