Docking phospholipase A2 on membranes using electrostatic potential-modulated spin relaxation magnetic resonance.
A method involving electron paramagnetic resonance spectroscopy of a site-selectively spin-labeled peripheral membrane protein in the presence and absence of membranes and of a water-soluble spin relaxant (chromium oxalate) has been developed to determine how bee venom phospholipase A2 sits on the membrane. Theory based on the Poisson-Boltzmann equation shows that the rate of spin relaxation of a protein-bound nitroxide by a membrane-impermeant spin relaxant depends on the distance (up to tens of angstroms) from the spin probe to the membrane. The measurements define the interfacial binding surface of this secreted phospholipase A2.
Lin
Y
Y
Department of Chemistry, Biochemistry, University of Washington, Box 351700, Seattle, WA 98195-1700, USA.
Nielsen
R
R
Murray
D
D
Hubbell
W L
WL
Mailer
C
C
Robinson
B H
BH
Gelb
M H
MH
D001514:Bee Venoms / Q000737:chemistry; D001665:Binding Sites; D002840:Chromates; D004578:Electron Spin Resonance Spectroscopy; D020404:Glycerophospholipids; D008081:Liposomes; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008567:Membranes, Artificial; D008958:Models, Molecular; D009154:Mutation; D010070:Oxalates; D010712:Phosphatidic Acids; D010741:Phospholipases A / Q000032:analysis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D054467:Phospholipases A2; D013113:Spin Labels; D013499:Surface Properties
D001514:Bee Venoms; D002840:Chromates; D020404:Glycerophospholipids; D008081:Liposomes; D008565:Membrane Proteins; D008567:Membranes, Artificial; D010070:Oxalates; D010712:Phosphatidic Acids; D013113:Spin Labels; C054506:tris(oxalato)chromate(III); C029313:dimyristoylmethylphosphatidic acid; D010741:Phospholipases A; D054467:Phospholipases A2