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Identification of a novel ATPase activity in 14‐3‐3 proteins – Evidence from enzyme kinetics, structure guided modeling and mutagenesis studies
Author(s) -
Ramteke Manoj P.,
Shelke Pradnya,
Ramamoorthy Vidhya,
Somavarapu Arun Kumar,
Gautam Amit Kumar Singh,
Nanaware Padma P.,
Karanam Sudheer,
Mukhopadhyay Sami,
Venkatraman Prasanna
Publication year - 2014
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2013.11.008
Subject(s) - mutagenesis , docking (animal) , gene isoform , homology modeling , dimer , chemistry , enzyme , biochemistry , binding site , recombinant dna , enzyme kinetics , plasma protein binding , biophysics , mutation , active site , biology , gene , medicine , nursing , organic chemistry
14‐3‐3 Proteins bind phosphorylated sequences in proteins and regulate multiple cellular functions. For the first time, we show that pure recombinant human 14‐3‐3 ζ, γ, ε and τ isofoms hydrolyze ATP with similar K m and k cat values. In sharp contrast the sigma isoform has no detectable activity. Docking studies identify two putative binding pockets in 14‐3‐3 zeta. Mutation of D124A in the amphipathic pocket enhances binding affinity and catalysis. Mutation of a critical Arg (R55A) at the dimer interface in zeta reduces binding and decreases catalysis. These experimental results coincide with a binding pose at the dimer interface. This newly identified function could be a moon lighting function in some of these isoforms.