Binding of polylysine to GroEL. Inhibition of the refolding of mMDH.
Luminescence techniques have been used to investigate the interaction of GroEL with polylysine tagged with a fluorescent probe. The fluorescence emitted by anthraniloyl-polylysine, upon excitation at 320 nm, is enhanced by the addition of stoichiometric amounts of GroEL. The equilibrium dissociation constant of the complex (Kd=50 nM) was determined by fluorometric titrations. The rate and extent of recovery of the catalytic activity of denatured mitochondrial malate dehydrogenase, assisted by GroEL, is influenced by either polylysine or anthraniloyl-polylysine. It is suggested that interaction of the positively charged poly-amino acid with the apical domain of GroEL prevents binding of the unfolded protein substrate.
Lau
C K
CK
Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, General Office, GH 602, Hung Hom, Kowloon, Hong Kong, China.
Churchich
J E
JE
D018834:Chaperonin 60 / Q000737:chemistry*; D004789:Enzyme Activation; D004926:Escherichia coli; D005456:Fluorescent Dyes; D008163:Luminescent Measurements; D008291:Malate Dehydrogenase / Q000737:chemistry*; D008928:Mitochondria / Q000201:enzymology; D009994:Osmolar Concentration; D011107:Polylysine / Q000737:chemistry*; D011489:Protein Denaturation; D017510:Protein Folding
D018834:Chaperonin 60; D005456:Fluorescent Dyes; D011107:Polylysine; D008291:Malate Dehydrogenase