title: Amyloid-type fiber formation in control of enzyme action: interfacial activation of phospholipase A2.
abstract: The lag-burst behavior in the action of phospholipase A(2) (PLA(2)) on 1,2-dipalmitoyl-sn-glycero-3-phosphocholine was investigated at temperatures slightly offset from the main phase transition temperature T(m) of this lipid, thus slowing down the kinetics of the activation process. Distinct stages leading to maximal activity were resolved using a combination of fluorescence parameters, including Förster resonance energy transfer between donor- and acceptor-labeled enzyme, fluorescence anisotropy, and lifetime, as well as thioflavin T fluorescence enhancement. We showed that the interfacial activation of PLA(2), evident after the preceding lag phase, coincides with the formation of oligomers staining with thioflavin T and subsequently with Congo red. Based on previous studies and our findings here, we propose a novel mechanism for the control of PLA(2), involving amyloid protofibrils with highly augmented enzymatic activity. Subsequently, these protofibrils form "mature" fibrils, devoid of activity. Accordingly, the process of amyloid formation is used as an on-off switch to obtain a transient burst in enzymatic catalysis.
authors.affiliation: Helsinki Biophysics and Biomembrane Group, Medical Biochemistry, Institute of Biomedicine, University of Helsinki, Finland.
authors.forename: Christian
authors.initials: C
authors.lastname: Code
authors.forename: Yegor
authors.initials: Y
authors.lastname: Domanov
authors.forename: Arimatti
authors.initials: A
authors.lastname: Jutila
authors.forename: Paavo K J
authors.initials: PK
authors.lastname: Kinnunen
mesh_terms: D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry*; D000682:Amyloid / Q000737:chemistry*; D003198:Computer Simulation; D003460:Crystallization / Q000379:methods*; D019281:Dimerization; D004789:Enzyme Activation; D008956:Models, Chemical; D008958:Models, Molecular; D054467:Phospholipases A2 / Q000737:chemistry*; D013499:Surface Properties
chemical_list: D000682:Amyloid; D015060:1,2-Dipalmitoylphosphatidylcholine; D054467:Phospholipases A2