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Titolo:
Analysis of the thyrotropin-releasing hormone-degrading ectoenzyme by site-directed mutagenesis of cysteine residues - Cys68 is involved in disulfide-linked dimerization
Autore:
Papadopoulos, T; Heuer, H; Bauer, K;
Indirizzi:
Max Planck Inst Expt Endokrinol, D-30603 Hannover, Germany Max Planck InstExpt Endokrinol Hannover Germany D-30603 nnover, Germany
Titolo Testata:
EUROPEAN JOURNAL OF BIOCHEMISTRY
fascicolo: 9, volume: 267, anno: 2000,
pagine: 2617 - 2623
SICI:
0014-2956(200005)267:9<2617:AOTTHE>2.0.ZU;2-Q
Fonte:
ISI
Lingua:
ENG
Soggetto:
GUINEA-PIG BRAIN; PYROGLUTAMATE AMINOPEPTIDASE; PRIMARY CULTURES; PITUITARY-GLAND; CLONING; SPECIFICITY; LOCALIZATION; PURIFICATION; THYROLIBERIN; PEPTIDASE;
Keywords:
metallopeptidase; Zn-dependent aminopeptidase; cysteine residues; covalent dimerization; site-directed mutagenesis;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
40
Recensione:
Indirizzi per estratti:
Indirizzo: Bauer, K Max Planck Inst Expt Endokrinol, POB 610309, D-30603 Hannover, Germany Max Planck Inst Expt Endokrinol POB 610309 Hannover Germany D-30603
Citazione:
T. Papadopoulos et al., "Analysis of the thyrotropin-releasing hormone-degrading ectoenzyme by site-directed mutagenesis of cysteine residues - Cys68 is involved in disulfide-linked dimerization", EUR J BIOCH, 267(9), 2000, pp. 2617-2623

Abstract

Thyrotropin-releasing hormone-degrading ectoenzyme is a member of the M1 family of Zn-dependent aminopeptidases and catalyzes the degradation of thyrotropin-releasing hormone (TRH; Glp-His-Pro-NH2). Cloning of the cDNA of this enzyme and biochemical studies revealed that the large extracellular domain of the enzyme with the catalytically active site contains nine cysteineresidues that are highly conserved among species. To investigate the functional role of these cysteines in TRH-DE we used a site-directed mutagenesisapproach and replaced individually each cysteine by a serine residue. The results revealed that the proteolytically truncated and enzymatically fullyactive enzyme consists of two identical subunits that are associated noncovalently by protein-protein interactions but not via interchain S-S bridges. The eight cysteines contained within this region are all important for the structure of the individual subunit and the enzymatic activity, which is dramatically reduced in all mutant enzymes. This is even true for the four cysteines that are clustered within the C-terminal domain remote from the Zn-binding consensus sequence HEICH. In contrast, Cys68, which resides within the stalk region seven residues from the end of the hydrophobic membrane-spanning domain, can be replaced by serine without a significant change in the enzymatic activity. Interestingly, this residue is involved in the formation of an interchain disulfide bridge. Covalent dimerization of the subunits, however, does not seem to be essential for efficient biosynthesis, enzymatic activity and trafficking to the cell surface.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 29/03/20 alle ore 18:17:03