A structural homologue of colipase in black mamba venom revealed by NMR floating disulphide bridge analysis.
The solution structure of mamba intestinal toxin 1 (MIT1), isolated from Dendroaspis polylepis polylepis venom, has been determined. This molecule is a cysteine-rich polypeptide exhibiting no recognised family membership. Resistance to MIT1 to classical specific endoproteases produced contradictory NMR and biochemical information concerning disulphide-bridge topology. We have used distance restraints allowing ambiguous partners between S atoms in combination with NMR-derived structural information, to correctly determine the disulphide-bridge topology. The resultant solution structure of MIT1, determined to a resolution of 0.5 A, reveals an unexpectedly similar global fold with respect to colipase, a protein involved in fatty acid digestion. Colipase exhibits an analogous resistance to endoprotease activity, indicating for the first time the possible topological origins of this biochemical property. The biochemical and structural homology permitted us to propose a mechanically related digestive function for MIT1 and provides novel information concerning snake venom protein evolution.
Boisbouvier
J
J
Institut de Biologie Structurale Jean-Pierre Ebel (CEA-CNRS), 41 Av. des Martyrs, 38027 Grenoble, France.
Albrand
J P
JP
Blackledge
M
M
Jaquinod
M
M
Schweitz
H
H
Lazdunski
M
M
Marion
D
D
D000818:Animals; D003089:Colipases / Q000737:chemistry*; D004220:Disulfides / Q000737:chemistry*; D004546:Elapid Venoms / Q000737:chemistry*; D017815:Elapidae; D010450:Endopeptidases / Q000378:metabolism; D019143:Evolution, Molecular; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010720:Phosphines; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D019163:Reducing Agents; D017386:Sequence Homology, Amino Acid; D013552:Swine
D003089:Colipases; D004220:Disulfides; D004546:Elapid Venoms; D010450:Endopeptidases; D010720:Phosphines; D019163:Reducing Agents; C080938:tris(2-carboxyethyl)phosphine