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Bacterially synthesized vertebrate calmodulin is a specific substrate for ubiquitination.

Publication ,  Journal Article
Gregori, L; Marriott, D; Putkey, JA; Means, AR; Chau, V
Published in: J Biol Chem
February 25, 1987

Calmodulin purified from bacteria which express a cloned chicken calmodulin gene can be selectively conjugated with ubiquitin, using enzymes present in reticulocyte extracts. Analyses of peptide products generated from limited proteolytic digestion of the calmodulin conjugate containing a single ubiquitin indicate that lysine 115 on calmodulin is the site of linkage. This linkage site is identical to that previously reported for calmodulin purified from Dictyostelium discoideum. Substrate-dependent ATP hydrolysis by a partially purified ubiquitin conjugation enzyme system from reticulocyte extracts was used to determine the enzyme affinity to calmodulin. Km values of 7 and 9 microM were determined for dictyostelium and the bacterially expressed calmodulin, respectively. The bacterially expressed calmodulin, unlike the Dictyostelium protein, can also form conjugates containing a 2-5 molar ratio of ubiquitin but at a slower rate than that observed for conjugation at lysine 115. Results from these studies further support our hypothesis that the post-translational methylation of lysine 115 found in most forms of calmodulin serves the important function of protecting calmodulin from ubiquitination and from degradation by the cytoplasmic ubiquitin-dependent proteolytic pathway. The capability of the bacterially expressed calmodulin to form conjugates with a high molar ratio of ubiquitin suggests that the post-translational acetylation of the N terminus of calmodulin may serve a similar function.

Duke Scholars

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

February 25, 1987

Volume

262

Issue

6

Start / End Page

2562 / 2567

Location

United States

Related Subject Headings

  • Ubiquitins
  • Kinetics
  • Electrophoresis, Polyacrylamide Gel
  • Dictyostelium
  • Cloning, Molecular
  • Chickens
  • Cattle
  • Calmodulin
  • Biochemistry & Molecular Biology
  • Animals
 

Citation

APA
Chicago
ICMJE
MLA
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Gregori, L., Marriott, D., Putkey, J. A., Means, A. R., & Chau, V. (1987). Bacterially synthesized vertebrate calmodulin is a specific substrate for ubiquitination. J Biol Chem, 262(6), 2562–2567.
Gregori, L., D. Marriott, J. A. Putkey, A. R. Means, and V. Chau. “Bacterially synthesized vertebrate calmodulin is a specific substrate for ubiquitination.J Biol Chem 262, no. 6 (February 25, 1987): 2562–67.
Gregori L, Marriott D, Putkey JA, Means AR, Chau V. Bacterially synthesized vertebrate calmodulin is a specific substrate for ubiquitination. J Biol Chem. 1987 Feb 25;262(6):2562–7.
Gregori, L., et al. “Bacterially synthesized vertebrate calmodulin is a specific substrate for ubiquitination.J Biol Chem, vol. 262, no. 6, Feb. 1987, pp. 2562–67.
Gregori L, Marriott D, Putkey JA, Means AR, Chau V. Bacterially synthesized vertebrate calmodulin is a specific substrate for ubiquitination. J Biol Chem. 1987 Feb 25;262(6):2562–2567.

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

February 25, 1987

Volume

262

Issue

6

Start / End Page

2562 / 2567

Location

United States

Related Subject Headings

  • Ubiquitins
  • Kinetics
  • Electrophoresis, Polyacrylamide Gel
  • Dictyostelium
  • Cloning, Molecular
  • Chickens
  • Cattle
  • Calmodulin
  • Biochemistry & Molecular Biology
  • Animals