title: Identification of a novel ATPase activity in 14-3-3 proteins--evidence from enzyme kinetics, structure guided modeling and mutagenesis studies.
abstract: 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 Km and kcat 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.
authors.affiliation: Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre (TMC), Kharghar, Navi Mumbai 410210, India.
authors.forename: Manoj P
authors.initials: MP
authors.lastname: Ramteke
authors.affiliation: Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre (TMC), Kharghar, Navi Mumbai 410210, India.
authors.forename: Pradnya
authors.initials: P
authors.lastname: Shelke
authors.affiliation: Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre (TMC), Kharghar, Navi Mumbai 410210, India.
authors.forename: Vidhya
authors.initials: V
authors.lastname: Ramamoorthy
authors.affiliation: Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre (TMC), Kharghar, Navi Mumbai 410210, India.
authors.forename: Arun Kumar
authors.initials: AK
authors.lastname: Somavarapu
authors.affiliation: Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre (TMC), Kharghar, Navi Mumbai 410210, India.
authors.forename: Amit Kumar Singh
authors.initials: AK
authors.lastname: Gautam
authors.affiliation: Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre (TMC), Kharghar, Navi Mumbai 410210, India.
authors.forename: Padma P
authors.initials: PP
authors.lastname: Nanaware
authors.affiliation: Vlife Sciences Technologies Pvt. Ltd., 2nd Floor, Plot No-05, Ram Indu Park, Baner Road, Pune 411045, India.
authors.forename: Sudheer
authors.initials: S
authors.lastname: Karanam
authors.affiliation: Vlife Sciences Technologies Pvt. Ltd., 2nd Floor, Plot No-05, Ram Indu Park, Baner Road, Pune 411045, India.
authors.forename: Sami
authors.initials: S
authors.lastname: Mukhopadhyay
authors.affiliation: Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre (TMC), Kharghar, Navi Mumbai 410210, India. Electronic address: vprasanna@actrec.gov.in.
authors.forename: Prasanna
authors.initials: P
authors.lastname: Venkatraman
mesh_terms: D048948:14-3-3 Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism*; D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D015394:Molecular Structure; D016296:Mutagenesis; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: D048948:14-3-3 Proteins; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases
keywords: 14-3-3; 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; AMP-PCP; ATP hydrolysis; ATP-γ-S; Chaperone; Ci; Docking; HEPES; MALDI; Mutation; Ni-NTA; PDB; PLP; RMSD; TOF; WT; adenosine 5′-(3-thiotriphosphate); adenylylmethylenediphosphonate; curie; matrix assisted laser desorption ionization; nickel-nitriloacetic acid; piecewise linear pairwise potential; protein data bank; root mean-square deviation; tandem time of flight; wild-type