X-ray structure of a bifunctional protein kinase in complex with its protein substrate HPr

被引:70
作者
Fieulaine, S
Morera, S
Poncet, S
Mijakovic, I
Galinier, A
Janin, J
Deutscher, J
Nessler, S [1 ]
机构
[1] CNRS, UPR 9063, Lab Enzymol & Biochim Struct, F-91198 Gif Sur Yvette, France
[2] CNRS, URA 1925, Lab Genet Microorganismes, Inst Natl Rech Agron, F-78850 Thiverval Grignon, France
[3] CNRS, UPR 9043, Chim Bacterienne Lab, F-13402 Marseille, France
关键词
D O I
10.1073/pnas.192368699
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
HPr kinase/phosphorylase (HprK/P) controls the phosphorylation state of the phosphocarrier protein HPr and regulates the utilization of carbon sources by Gram-positive bacteria. It catalyzes both the ATP-dependent phosphorylation of Ser-46 of HPr and its dephosphorylation by phosphorolysis. The latter reaction uses inorganic phosphate as substrate and produces pyrophosphate. We present here two crystal structures of a complex of the catalytic domain of Lactobacillus casei HprK/P with Bacillus subtilis HPr, both at 2.8-Angstrom resolution. One of the structures was obtained in the presence of excess pyrophosphate, reversing the phosphorolysis reaction and contains serine-phosphorylated HPr. The complex has six HPr molecules bound to the hexameric kinase. Two adjacent enzyme subunits are in contact with each HPr molecule, one through its active site and the other through its C-terminal helix. In the complex with serine-phosphorylated HPr, a phosphate ion is in a position to perform a nucleophilic attack on the phosphoserine. Although the mechanism of the phosphorylation reaction resembles that of eukaryotic protein kinases, the dephosphorylation by inorganic phosphate is unique to the HprK/P family of kinases. This study provides the structure of a protein kinase in complex with its protein substrate, giving insights into the chemistry of the phospho-transfer reactions in both directions.
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页码:13437 / 13441
页数:5
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