Spectroscopic characterization of the interaction between calmodulin-dependent protein kinase I and calmodulin

Gomes, Aldrin V; Barnes, Junor A; Vogel, Hans J

Gomes, Aldrin V; Barnes, Junor A; Vogel, Hans J.

Afiliação

Gomes, Aldrin V; University of Calgary. Department of Biological Sciences. Alberta. Canada
Barnes, Junor A; The University of the West Indies. Department of Preclinical Sciences. Biochemistry Unit. St. Augustine. Trinidad and Tobago
Vogel, Hans J; University of Calgary. Department of Biological Sciences. Alberta. Canada

Archives of biochemistry and biophysics ; 379(1): 28-36, July 2000. ilus, gra

Article em En | MedCarib | ID: med-17303

Biblioteca responsável: TT5

Localização: TT5; W1 AR449

ABSTRACT

ABSTRACT

Calmodulin-dependent protein kinase I (CaM kinase I) is a member of the expanding class of protein kinases that are regulated by calmodulin (CaM). Its putative CaM -binding region is believed to occur within a 22-residue sequence (amino acids 299-320). This sequence was chemically synthesized and utilized for CaM interaction studies. GEl band shift assays and densitometry experiments with intact CAM kinase I and the CAM-binding domain peptide (CaMKIp) reveal that they bind in an analogous manner, giving rise to 11 complexes. Fluorescence analysis using dansyl-CaM showed that confirmational changes CaM on binding CaM kinase I orCaMKIp were nearly identical, suggesting that the peptide mimicked the CaM-binding ability of the intact protein. In the presence of Ca, the peptide displays an enhancement of its unique Trp fluorescence as well as a marked blue shift of the emission maximum, reflecting a transfer to a more rigid, less polar environment. Quenching studies, using acrylamide, confirmed that the Trp in the peptide on binding CaM is no longer freely exposed to solvent as is the case for the free peptide. Studies with a series of MET small mutants of CaM showed that the Trp-containing N-terminal lobe of CaM was bound to the C-terminal lobe of CaM. Near-UV CD spectra also indicated that the Trp of the peptide and Phe residues of the protein are involved in the binding. These results show that the CaM-binding domain of CaM kinase I binds to CaM in a manner analogous to that of myosin light chain kinase (AU)

Assuntos

Proteínas Quinases Dependentes de Cálcio-Calmodulina; Calmodulina; Peptídeos; Dicroísmo Circular

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Coleções: 01-internacional Base de dados: MedCarib Assunto principal: Peptídeos / Calmodulina / Dicroísmo Circular / Proteínas Quinases Dependentes de Cálcio-Calmodulina Idioma: En Revista: Archives of biochemistry and biophysics Ano de publicação: 2000 Tipo de documento: Article

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