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Nucleoside diphosphate kinase from bovine retina: purification, subcellular localization, molecular cloning, and three-dimensional structure.

Publication ,  Journal Article
Abdulaev, NG; Karaschuk, GN; Ladner, JE; Kakuev, DL; Yakhyaev, AV; Tordova, M; Gaidarov, IO; Popov, VI; Fujiwara, JH; Chinchilla, D; Ridge, KD ...
Published in: Biochemistry
October 1998

The biochemical and structural properties of bovine retinal nucleoside diphosphate kinase were investigated. The enzyme showed two polypeptides of approximately 17.5 and 18.5 kDa on SDS-PAGE, while isoelectric focusing revealed seven to eight proteins with a pI range of 7.4-8.2. Sedimentation equilibrium yielded a molecular mass of 96 +/- 2 kDa for the enzyme. Carbohydrate analysis revealed that both polypeptides contained Gal, Man, GlcNAc, Fuc, and GalNac saccharides. Like other nucleoside diphosphate kinases, the retinal enzyme showed substantial differences in the Km values for various di- and triphosphate nucleotides. Immunogold labeling of bovine retina revealed that the enzyme is localized on both the membranes and in the cytoplasm. Screening of a retinal cDNA library yielded full-length clones encoding two distinct isoforms (NBR-A and NBR-B). Both isoforms were overexpressed in Escherichia coli and their biochemical properties compared with retinal NDP-kinase. The structures of NBR-A and NBR-B were determined by X-ray crystallography in the presence of guanine nucleotide(s). Both isoforms are hexameric, and the fold of the monomer is similar to other nucleoside diphosphate kinase structures. The NBR-A active site contained both a cGMP and a GDP molecule each bound at half occupancy while the NBR-B active site contained only cGMP.

Duke Scholars

Published In

Biochemistry

ISSN

0006-2960

Publication Date

October 1998

Volume

37

Issue

40

Start / End Page

13958 / 13967

Location

united states

Related Subject Headings

  • Biochemistry & Molecular Biology
  • 1101 Medical Biochemistry and Metabolomics
  • 0601 Biochemistry and Cell Biology
  • 0304 Medicinal and Biomolecular Chemistry
 

Citation

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Abdulaev, N. G., Karaschuk, G. N., Ladner, J. E., Kakuev, D. L., Yakhyaev, A. V., Tordova, M., … Ridge, K. D. (1998). Nucleoside diphosphate kinase from bovine retina: purification, subcellular localization, molecular cloning, and three-dimensional structure. Biochemistry, 37(40), 13958–13967.
Abdulaev, N. G., G. N. Karaschuk, J. E. Ladner, D. L. Kakuev, A. V. Yakhyaev, M. Tordova, I. O. Gaidarov, et al. “Nucleoside diphosphate kinase from bovine retina: purification, subcellular localization, molecular cloning, and three-dimensional structure.Biochemistry 37, no. 40 (October 1998): 13958–67.
Abdulaev NG, Karaschuk GN, Ladner JE, Kakuev DL, Yakhyaev AV, Tordova M, et al. Nucleoside diphosphate kinase from bovine retina: purification, subcellular localization, molecular cloning, and three-dimensional structure. Biochemistry. 1998 Oct;37(40):13958–67.
Abdulaev NG, Karaschuk GN, Ladner JE, Kakuev DL, Yakhyaev AV, Tordova M, Gaidarov IO, Popov VI, Fujiwara JH, Chinchilla D, Eisenstein E, Gilliland GL, Ridge KD. Nucleoside diphosphate kinase from bovine retina: purification, subcellular localization, molecular cloning, and three-dimensional structure. Biochemistry. 1998 Oct;37(40):13958–13967.
Journal cover image

Published In

Biochemistry

ISSN

0006-2960

Publication Date

October 1998

Volume

37

Issue

40

Start / End Page

13958 / 13967

Location

united states

Related Subject Headings

  • Biochemistry & Molecular Biology
  • 1101 Medical Biochemistry and Metabolomics
  • 0601 Biochemistry and Cell Biology
  • 0304 Medicinal and Biomolecular Chemistry