Substrate Recognition Mechanism of Carboxypeptidase Y

Nakase, Hiroshi; Murata, Shoichi; Ueno, Hiroshi

doi:10.1271/bbb.65.2465

Cited by 11 publications

(11 citation statements)

References 2 publications

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“…Two disulfide bonds, Cys193-Cys207 and Cys262-Cys268, are located at the beginning and ending of the V-shape helix, just like hinges. subsites, respectively, play critical roles (5,(18)(19)(20)(21)(22)(23)(24). Interestingly, a part of V-shape helix constru cts the S3 and S5 subsites.…”

Section: Serine-type Protease Of Yeast Saccharomycesmentioning

confidence: 99%

Amino acid substitution reveals the role of V-shape helix on construction of yeast carboxypeptidase Y

et al. 2014

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Carboxypeptidase Y (CPY) is a vacuolar serine-type protease of yeast Saccharomyces cerevisiae to release wide variety of L-amino acid including proline from C termini of peptides and proteins. According to three-dimensional structure of mature CPY determined crystallographically, there is a characteristic region constructed with two adjacent α-helices, called 'V-shape helix', which lies over the active site cavity just like a door. In order to clarify the role of V-shape helix, we attempted to immobilize the V-shape helix by introducing amino acid substitution to create additional disulfide bridge between the V-shape helix and the main body of CPY. Resulting mutant enzymes presented with misfolding or hydrolase activity loss, suggesting that replacement of the residues at or near V-shape helix would influence on the protein folding and enzyme activity. The results of this report demonstrated that the insertion domain including V-shape helix in α/β hydrolasefold serine carboxypeptidases plays a role in the construction of functional enzyme.

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Section: Serine-type Protease Of Yeast Saccharomycesmentioning

confidence: 99%

Amino acid substitution reveals the role of V-shape helix on construction of yeast carboxypeptidase Y

et al. 2014

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“…The 372delD LAMP‐2A mutation implies the modification of the residue that corresponds to the P5 substrate position (Schechter and Berger nomenclature) and to a certain extent might affect the HPP‐mediated proteolysis. This conclusion can be made after knowing that the affinity for substrates by carboxypeptidase Y, an HPP homologue, is contributed by a set of residues that also includes the one in P5 position [Nakase et al, 2001]. To comprehend the effects of the mutation on the LAMP‐2A/HPP interaction, we built the structural model of the DDDNF peptide (LAMP‐2A P 5 ‐P 1 residues) bound to HPP (model available upon request).…”

Section: To the Editormentioning

confidence: 99%

Molecular analysis of PRKAG2, LAMP2, and NKX2‐5 genes in a cohort of 125 patients with accessory atrioventricular connection

Esposito

Grutter

Drago

et al. 2009

American J of Med Genetics Pt A

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“…Тhe nonapeptide LNGGP GCSS (76-84), the sequence surrounding the active site serine FHIAG ESYAG HYIP (163-176) and the motif ICNWL GN (368-374) all showed significant conservation. Each of these motifs comprises components (underlined residues) of the substrate binding subsites, the oxyanion hole or the hydrogen bond network (Liao and Remington 1990;Nasr et al 1994;Jung et al 1999;Nakase et al 2001).…”

Section: Resultsmentioning

confidence: 99%

The Kluyveromyces lactis CPY homologous genes — Cloning and characterization of the KlPCL1 gene

et al. 2008

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A 3.85-kb genomic fragment containing the KlPCL1 gene, with an open reading frame (ORF) of 1359 bp, was isolated from Kluyveromyces lactis genomic library by heterologous colony hybridization using the Saccharomyces cerevisiae PRC1 (ScPRC1) gene as a probe. The KlPCL1 nucleotide sequence was identical to the KLLAOC17490g ORF of K. lactis and showed >55 % identity with S. cerevisiae YBR139w and PRC1 genes encoding carboxypeptidases. The deduced KlPcl1p amino acid sequence displayed strong similarities to yeast and higher eukaryotic carboxypeptidases. In silico analyses revealed that KlPcl1p contained several highly conserved regions characteristic of the serine-type carboxypeptidases, such as the catalytic triad in the active site and the LNGGPGCSS, FHIAGESYAGHYIP and ICNWLGN motifs involved in the substrate binding. All this suggests that the KlPCL1 gene product belongs to the serine carboxypeptidase family. Sporulation and ascus dissection of a diploid strain heterozygous for single-copy disruption of KlPCL1 revealed that this gene is not essential in K. lactis. Further analyses of haploid and diploid deletion mutants demonstrated that disruption of the KlPCL1 gene neither impaired sporulation nor affected growth abilities of K. lactis cells under a variety of physiological conditions, e.g., growth on different carbon sources, at various temperatures or pH of the medium, and under nitrogen depletion.

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Substrate Recognition Mechanism of Carboxypeptidase Y

Cited by 11 publications

References 2 publications

Amino acid substitution reveals the role of V-shape helix on construction of yeast carboxypeptidase Y

Amino acid substitution reveals the role of V-shape helix on construction of yeast carboxypeptidase Y

Molecular analysis of PRKAG2, LAMP2, and NKX2‐5 genes in a cohort of 125 patients with accessory atrioventricular connection

The Kluyveromyces lactis CPY homologous genes — Cloning and characterization of the KlPCL1 gene

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