PDBsum entry: 1kaz

Hydrolase

PDB id: 1kaz

Contents

Protein chain

378 a.a. *

Ligands

ATP

Metals

_MG

__K ×3

_CL ×2

Waters ×429

* Residue conservation analysis

PDB id:

1kaz

Name:

Hydrolase

Title:

70kd heat shock cognate protein atpase domain, k71e mutant

Structure:

70kd heat shock cognate protein. Chain: a. Fragment: atpase domain. Synonym: hsc70. Engineered: yes. Mutation: yes

Source:

Bos taurus. Cattle. Organism_taxid: 9913

Resolution:

1.70Å R-factor: 0.202 R-free: 0.234

Authors:

M.C.O'Brien,K.M.Flaherty,D.B.Mckay

Key ref:

M.C.O'Brien et al. (1996). Lysine 71 of the chaperone protein Hsc70 Is essential for ATP hydrolysis. J Biol Chem, 271, 15874-15878. PubMed id: 8663302 DOI: 10.1074/jbc.271.27.15874

Date:

15-Apr-96 Release date: 08-Nov-96

Protein chain

P19120  (HSP7C_BOVIN) -  Heat shock cognate 71 kDa protein

Seq: 650 a.a.

Struc: 378 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

 Gene Ontology (GO) functional annotation 

• Biochemical function: ATP binding (1 term)

DOI no: 10.1074/jbc.271.27.15874

J Biol Chem 271:15874-15878 (1996)

PubMed id: 8663302

Lysine 71 of the chaperone protein Hsc70 Is essential for ATP hydrolysis.

M.C.O'Brien, K.M.Flaherty, D.B.McKay.

ABSTRACT

It has been proposed that lysine 71 of the bovine 70-kDa heat shock cognate protein might participate in catalysis of ATP hydrolysis by stabilizing an H2O molecule or an OH- ion for nucleophilic attack on the gamma-phosphate of the nucleotide (Flaherty, K. M., Wilbanks, S. M., DeLuca-Flaherty, C., and McKay, D. B. (1994) J. Biol. Chem. 12899-12907; Wilbanks, S. M., DeLuca-Flaherty, C., and McKay, D. B. (1994) J. Biol. Chem. 269, 12893-12898). To test this hypothesis, lysine 71 of the ATPase fragment 70-kDa heat shock cognate protein has been mutated to glutamic acid, methionine, and alanine; and the kinetic and structural properties of the mutant proteins have been determined. All three mutant proteins are devoid of measurable ATP hydrolysis activity. Crystal structures of the mutant proteins have been determined to a resolution of 1.7 A; all three have ATP in the nucleotide binding site. These data identify lysine 71 as a residue that is essential for chemical hydrolysis of ATP.

Selected figure(s)

Figure 1.
Fig. 1. Anomalous difference Fourier map for the K71E structure. Contour level, 2.0 . In the stick model, oxygen atoms are shown in black, metal ions are shown as heavy crosses, and other atoms are gray. Three large peaks are due to K+ ions; three smaller peaks are due to phosphorus atoms. The side chain of residue 71 is labeled K71E.

Figure 2.
Fig. 2. Structures of wild-type and mutant proteins in the triphosphate binding region of the ATPase fragment. Phosphorus atoms are black, carbon atoms are white, and others are gray. Metal ions are labeled. Subsets of the interactions with Mg2+ and K+ are shown by dotted lines. A, K71E; B, K71M; C, K71A; D, active site region in the wild-type protein with ADP and P[i] bound.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (1996, 271, 15874-15878) copyright 1996.