Mutagenesis of residues involved in control of the Ca2+ entry pathway and conformational changes associated with Ca2+ binding in the SR Ca2+-ATPase

E(1)Ca(2) transition) in the wild type and mutants of the sarcoplasmic reticulum Ca(2+)-ATPase expressed in mammalian cells. Cluster mutations as well as single mutations in transmembrane segment M3 resulted in conspicuous effects on the rate of Ca(2+) migration. Furthermore, mutation of Asp(59) in transmembrane segment M1 to arginine exerted a profound effect on Ca(2+) interaction. The data demonstrate an important role for M3 residues in control of the Ca(2+) entry pathway and provide functional evidence in support of a close relationship between this pathway and the water-accessible channel leading between transmembrane segments M1 and M3 in the thapsigargin stabilized E(2) structure. In addition, rapid kinetic measurements demonstrated that the hydrogen bond network involving Asp(813) of loop L6-7 and Lys(758) of M5 is important for the E(2) --> E(1)Ca(2) transition. -->

ISSN:	0077-8923
Přístupová URL adresa:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::833b6d52732ba1963224160d7e5d218a https://pubmed.ncbi.nlm.nih.gov/12763777
Rights:	CLOSED
Přírůstkové číslo:	edsair.doi.dedup.....833b6d52732ba1963224160d7e5d218a
Autor:	Anja P. Einholm, Bente Vilsen, Jens Peter Andersen, Johannes D. Clausen
Rok vydání:	2003
Předmět:	Models Molecular Thapsigargin Arginine Stereochemistry Protein Conformation ATPase Mutant Calcium-Transporting ATPases General Biochemistry Genetics and Molecular Biology Protein Structure Secondary Sarcoplasmic Reticulum Calcium-Transporting ATPases chemistry.chemical_compound History and Philosophy of Science Animals Point Mutation Amino Acid Sequence Phosphorylation Binding Sites biology Chemistry General Neuroscience Endoplasmic reticulum Wild type Transmembrane protein Peptide Fragments Recombinant Proteins Transmembrane domain Kinetics biology.protein Mutagenesis Site-Directed Calcium
Zdroj:	Annals of the New York Academy of Sciences. 986
ISSN:	0077-8923
Popis:	Rapid kinetic measurements were used to study the rate of Ca(2+) dissociation from the high-affinity Ca(2+) sites of the dephosphoenzyme (i.e., from the E(1)Ca(2) form toward the cytoplasmic side) as well as the rate of Ca(2+) binding with associated conformational changes (E(2) --> E(1)Ca(2) transition) in the wild type and mutants of the sarcoplasmic reticulum Ca(2+)-ATPase expressed in mammalian cells. Cluster mutations as well as single mutations in transmembrane segment M3 resulted in conspicuous effects on the rate of Ca(2+) migration. Furthermore, mutation of Asp(59) in transmembrane segment M1 to arginine exerted a profound effect on Ca(2+) interaction. The data demonstrate an important role for M3 residues in control of the Ca(2+) entry pathway and provide functional evidence in support of a close relationship between this pathway and the water-accessible channel leading between transmembrane segments M1 and M3 in the thapsigargin stabilized E(2) structure. In addition, rapid kinetic measurements demonstrated that the hydrogen bond network involving Asp(813) of loop L6-7 and Lys(758) of M5 is important for the E(2) --> E(1)Ca(2) transition.
Databáze:	OpenAIRE
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