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Purification and characterization of nucleoside diphosphate kinase from the brain of Bombyx mori
Author(s) -
Uno Tomohide,
Ueno Mayumi,
Kikuchi Michiko,
Aizono Yasuo
Publication year - 2002
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.10037
Subject(s) - biochemistry , biology , kinase , mitogen activated protein kinase kinase , map2k7 , dephosphorylation , microbiology and biotechnology , protein kinase a , phosphatase , phosphorylation , cyclin dependent kinase 2 , enzyme , cyclin dependent kinase 9
Nucleoside diphosphate kinase in the brain of Bombyx mori was purified by ammonium sulfate fractionation, and a sequence of chromatographies on DEAE‐Cellulofine, hydroxyapatite, Mono‐S, and Mono‐Q column. The purified enzyme preparation was found to be electrophoretically homogeneous on SDS‐PAGE, and its molecular mass was determined to be 18 kDa. The purified protein was digested and the amino acid sequences of resulting peptides were determined. The enzyme showed high similarity to the amino acid sequences of the Drosophila NDP kinase. The enzyme showed NDP kinase activity and mediated the phosphorylation of myelin basic protein. Gel filtration and Hill plot analysis indicate that the purified NDP kinase forms a tetramer and shows little interaction among substrates. Dephosphorylation of NDP kinase by bacterial alkaline phosphatase increased NDP kinase activity. This result indicates that phosphorylation of NDP kinase represses NDP kinase activity. Arch. Insect Biochem. Physiol. 50:147–155, 2002. © 2002 Wiley‐Liss, Inc.