Identification of an autoinhibitory domain in calcineurin

Yoshiaki Hashimoto, Brian A. Perrino, Thomas Soderling

Research output: Contribution to journalArticle

214 Citations (Scopus)

Abstract

The hypothesis that calcineurin, the Ca2+/calmodulin-dependent protein phosphatase, contains an autoinhibitory domain was tested using synthetic peptides corresponding to regions of the carboxyl-terminus of calcineurin. Of the several peptides analyzed, one, containing residues I-T-S-F-E-E-A-K-G-L-D-R-I-N-E-R-M-P-P-R-R-D-A-M-P, gave complete inhibition of its protein phosphatase activity. Using [32P]myosin light chain as substrate an IC50 of about 10 μM was obtained with either native calcineurin, assayed in the presence of Ca2+/calmodulin, or with calcineurin subjected to partial proteolysis which converts it to a fully active phosphatase when assayed in the presence of [ethylenebis(oxyethylenenitrilo)]tetraacetic acid. With 50 mM p-nitrophenylphosphate as substrate an IC50 of about 40 μM was observed. Studies with overlapping peptides suggested that the sequence P-P-R-R-D-A-M-P was essential but not sufficient for the observed inhibition. Kinetic analysis indicated that the inhibition of phosphatase activity was not competitive with respect to [32P]myosin light chain. This peptide did not show significant inhibition of the catalytic subunits of protein phosphatases type I or type IIA or of Ca2+/ calmodulin-dependent protein kinase II. These results indicate that amino acids within this sequence of calcineurin constitute a unique autoinhibitory domain which interacts with the active site and is responsible for the low basal phosphatase activity in the absence of Ca2+/calmodulin.

Original languageEnglish (US)
Pages (from-to)1924-1927
Number of pages4
JournalJournal of Biological Chemistry
Volume265
Issue number4
StatePublished - Feb 5 1990
Externally publishedYes

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Calcineurin
Phosphoprotein Phosphatases
Calmodulin
Phosphoric Monoester Hydrolases
Myosin Light Chains
Peptides
Inhibitory Concentration 50
Catalytic Domain
Proteolysis
Calcium-Calmodulin-Dependent Protein Kinase Type 2
Substrates
Amino Acid Sequence
Amino Acids
Kinetics
Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Hashimoto, Y., Perrino, B. A., & Soderling, T. (1990). Identification of an autoinhibitory domain in calcineurin. Journal of Biological Chemistry, 265(4), 1924-1927.

Identification of an autoinhibitory domain in calcineurin. / Hashimoto, Yoshiaki; Perrino, Brian A.; Soderling, Thomas.

In: Journal of Biological Chemistry, Vol. 265, No. 4, 05.02.1990, p. 1924-1927.

Research output: Contribution to journalArticle

Hashimoto, Y, Perrino, BA & Soderling, T 1990, 'Identification of an autoinhibitory domain in calcineurin', Journal of Biological Chemistry, vol. 265, no. 4, pp. 1924-1927.
Hashimoto Y, Perrino BA, Soderling T. Identification of an autoinhibitory domain in calcineurin. Journal of Biological Chemistry. 1990 Feb 5;265(4):1924-1927.
Hashimoto, Yoshiaki ; Perrino, Brian A. ; Soderling, Thomas. / Identification of an autoinhibitory domain in calcineurin. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 4. pp. 1924-1927.
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