Expression in Escherichia coli and characterization of the heat-stable inhibitor of the cAMP-dependent protein kinase

James Thomas, Scott M. Van Patten, Paul Howard, Kathleen H. Day, Ryan D. Mitchell, Tobin Sosnick, Jill Trewhella, Donal A. Walsh, Richard Maurer

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Pure heat-stable inhibitor of the cAMP-dependent protein kinase (PKI) has been isolated in high yield by using a bacterial expression vector constructed to synthesize the complete sequence of the rabbit muscle protein kinase inhibitor, plus an amino-terminal initiator methionine and glycine. Bacterially expressed PKI has an inhibitory activity identical to that of the protein isolated from rabbit skeletal muscle and, by gel filtration and gel electrophoresis, has the same physicochemical characteristics as the native physiological form of PKI. Fourier transformed infrared spectroscopy and CD establish that PKI has unusually large amounts of random coil and turn structures, with significantly smaller amounts of α-helix and β structures.

Original languageEnglish (US)
Pages (from-to)10906-10911
Number of pages6
JournalJournal of Biological Chemistry
Volume266
Issue number17
StatePublished - 1991
Externally publishedYes

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Cyclic AMP-Dependent Protein Kinases
Protein Kinases
Escherichia coli
Hot Temperature
Gels
Rabbits
Muscle Proteins
Protein Kinase Inhibitors
Electrophoresis
Methionine
Glycine
Gel Chromatography
Muscle
Infrared spectroscopy
Spectrum Analysis
Skeletal Muscle
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Thomas, J., Van Patten, S. M., Howard, P., Day, K. H., Mitchell, R. D., Sosnick, T., ... Maurer, R. (1991). Expression in Escherichia coli and characterization of the heat-stable inhibitor of the cAMP-dependent protein kinase. Journal of Biological Chemistry, 266(17), 10906-10911.

Expression in Escherichia coli and characterization of the heat-stable inhibitor of the cAMP-dependent protein kinase. / Thomas, James; Van Patten, Scott M.; Howard, Paul; Day, Kathleen H.; Mitchell, Ryan D.; Sosnick, Tobin; Trewhella, Jill; Walsh, Donal A.; Maurer, Richard.

In: Journal of Biological Chemistry, Vol. 266, No. 17, 1991, p. 10906-10911.

Research output: Contribution to journalArticle

Thomas, J, Van Patten, SM, Howard, P, Day, KH, Mitchell, RD, Sosnick, T, Trewhella, J, Walsh, DA & Maurer, R 1991, 'Expression in Escherichia coli and characterization of the heat-stable inhibitor of the cAMP-dependent protein kinase', Journal of Biological Chemistry, vol. 266, no. 17, pp. 10906-10911.
Thomas J, Van Patten SM, Howard P, Day KH, Mitchell RD, Sosnick T et al. Expression in Escherichia coli and characterization of the heat-stable inhibitor of the cAMP-dependent protein kinase. Journal of Biological Chemistry. 1991;266(17):10906-10911.
Thomas, James ; Van Patten, Scott M. ; Howard, Paul ; Day, Kathleen H. ; Mitchell, Ryan D. ; Sosnick, Tobin ; Trewhella, Jill ; Walsh, Donal A. ; Maurer, Richard. / Expression in Escherichia coli and characterization of the heat-stable inhibitor of the cAMP-dependent protein kinase. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 17. pp. 10906-10911.
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