Allosteric inhibition of the neuropeptidase neurolysin.
Neuropeptidases specialize in the hydrolysis of the small bioactive peptides that play a variety of signaling roles in the nervous and endocrine systems. One neuropeptidase, neurolysin, helps control the levels of the dopaminergic circuit modulator neurotensin and is a member of a fold group that includes the antihypertensive target angiotensin converting enzyme. We report the discovery of a potent inhibitor that, unexpectedly, binds away from the enzyme catalytic site. The location of the bound inhibitor suggests it disrupts activity by preventing a hinge-like motion associated with substrate binding and catalysis. In support of this model, the inhibition kinetics are mixed, with both noncompetitive and competitive components, and fluorescence polarization shows directly that the inhibitor reverses a substrate-associated conformational change. This new type of inhibition may have widespread utility in targeting neuropeptidases.
From the Department of Molecular and Cellular Biochemistry and the Center for Structural Biology, University of Kentucky, Lexington, Kentucky 40536.
Christina S
CS
Hines
From the Department of Molecular and Cellular Biochemistry and the Center for Structural Biology, University of Kentucky, Lexington, Kentucky 40536.
Kallol
K
Ray
From the Department of Molecular and Cellular Biochemistry and the Center for Structural Biology, University of Kentucky, Lexington, Kentucky 40536.
Jack J
JJ
Schmidt
From the Department of Molecular and Cellular Biochemistry and the Center for Structural Biology, University of Kentucky, Lexington, Kentucky 40536.
Fei
F
Xiong
Abbott Healthcare Products B.V., C.J. van Houtenlaan 36, 1381 CP, Weesp, The Netherlands, and.
Rolf W
RW
Feenstra
Abbott Healthcare Products B.V., C.J. van Houtenlaan 36, 1381 CP, Weesp, The Netherlands, and.
Mia
M
Pras-Raves
Abbott Healthcare Products B.V., C.J. van Houtenlaan 36, 1381 CP, Weesp, The Netherlands, and.
Jan Peter
JP
de Moes
Abbott Healthcare Products B.V., C.J. van Houtenlaan 36, 1381 CP, Weesp, The Netherlands, and.
Jos H M
JH
Lange
From the Department of Molecular and Cellular Biochemistry and the Center for Structural Biology, University of Kentucky, Lexington, Kentucky 40536.
Manana
M
Melikishvili
From the Department of Molecular and Cellular Biochemistry and the Center for Structural Biology, University of Kentucky, Lexington, Kentucky 40536.
Michael G
MG
Fried
Evotec (UK) Ltd., 114 Milton Park, Abingdon, OX14 4SA Oxfordshire, United Kingdom.
Paul
P
Mortenson
Evotec (UK) Ltd., 114 Milton Park, Abingdon, OX14 4SA Oxfordshire, United Kingdom.
Michael
M
Charlton
Evotec (UK) Ltd., 114 Milton Park, Abingdon, OX14 4SA Oxfordshire, United Kingdom.
Yogendra
Y
Patel
Evotec (UK) Ltd., 114 Milton Park, Abingdon, OX14 4SA Oxfordshire, United Kingdom.
Stephen M
SM
Courtney
Abbott Healthcare Products B.V., C.J. van Houtenlaan 36, 1381 CP, Weesp, The Netherlands, and.
Chris G
CG
Kruse
From the Department of Molecular and Cellular Biochemistry and the Center for Structural Biology, University of Kentucky, Lexington, Kentucky 40536, david.rodgers@uky.edu.
David W
DW
Rodgers
D000494:Allosteric Regulation; D000495:Allosteric Site; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D055162:Biocatalysis / Q000187:drug effects; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D005454:Fluorescence Polarization; D007700:Kinetics; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008956:Models, Chemical; D008958:Models, Molecular; D015394:Molecular Structure; D020125:Mutation, Missense; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D013379:Substrate Specificity
D004791:Enzyme Inhibitors; D008666:Metalloendopeptidases; C062316:neurolysin
Allosteric Regulation; Hydrolase; Metalloprotease; Neurochemistry; Neuropeptide; Peptidase; X-ray Crystallography