• 제목/요약/키워드: carboxypeptidases

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Isolation and Characterization of Four Carboxypeptidases in Canavalia lineata Cotyledons

  • Yang, Jong-Moon;Rhew, Tae-Hyong;Koh, Suck-Chan;Kwon, Young-Myung
    • BMB Reports
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    • 제28권5호
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    • pp.451-457
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    • 1995
  • Four carboxypeptidases, CP1, CP2, CP3, and CP4 were isolated from the cotyledons of germinating seedlings of Canavalia lineata by sequential chromatography on the following four columns: 1) CM-cellulose, 2) Sephacryl 5-300, 3) Procion red dye, and 4) Sephacryl S-200. A number of properties of the enzymes, such as substrate specificity, molecular weight, optimum pH, thermal stability, have been determined. Enzyme activities were measured using the Cbz(carbobenzoxy)-dipeptides containing phenylalanine at the penultimate position. The $K_m$ values of four carboxypeptidases for Cbz-Phe-Ala were 0.50, 0.65, 1.30, and 1.35 mM, respectively. The inhibition studies indicated that the four carboxypeptidases were all serine type. Each of the carboxypeptidases with molecular weights of 145, 114, 105, and 104 kDa, respectively, had the optimum enzyme activity at pH 5.0~6.0. And they were sensitive to high temperature.

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Dipeptidyl Carboxypeptidases에 의한 S-Hippuryl Thioglycolyl Glycine의 가수분해 (S-Hippuryl Thioglycolyl Glycine : A New Chromogenic Substrate for Dipeptidyl Carboxypeptidases)

  • 이현재
    • 대한화학회지
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    • 제19권4호
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    • pp.246-251
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    • 1975
  • Dideptidyl carboxypeptidases와 angiotenisn-coverting enzyme의 새로운 기질불질로서 thiol ester 인 S-Hippuryl thioglycolyl glycine을 합성하였으며, 이 기질에 의한 간편하고도 예민한 효소 활성도의 정량방법을 제시하였다. 이 경우 효소반응 생성물인 thioglycolyl glycine은 반응계중에 첨가한 5,5-dithio-bis-(2-nitrobenzoic acid), DTNB와 쉽게 반응하여 410nm에서 강한 흡광스펙트럼을 갖는 5-thio-2-nitrobenzoic acid(${\varepsilon}M=1.36{\times}10^4$)을 형성함으로서 효소의 새로운 미량정량 방법으로 이용 가치가 크다고 본다.

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HIF-1α-Dependent Induction of Carboxypeptidase A4 and Carboxypeptidase E in Hypoxic Human Adipose-Derived Stem Cells

  • Moon, Yunwon;Moon, Ramhee;Roh, Hyunsoo;Chang, Soojeong;Lee, Seongyeol;Park, Hyunsung
    • Molecules and Cells
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    • 제43권11호
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    • pp.945-952
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    • 2020
  • Hypoxia induces the expression of several genes through the activation of a master transcription factor, hypoxia-inducible factor (HIF)-1α. This study shows that hypoxia strongly induced the expression of two carboxypeptidases (CP), CPA4 and CPE, in an HIF-1α-dependent manner. The hypoxic induction of CPA4 and CPE gene was accompanied by the recruitment of HIF-1α and upregulation in the active histone modification, H3K4me3, at their promoter regions. The hypoxic responsiveness of CPA4 and CPE genes was observed in human adipocytes, human adipose-derived stem cells, and human primary fibroblasts but not mouse primary adipocyte progenitor cells. CPA4 and CPE have been identified as secreted exopeptidases that degrade and process other secreted proteins and matrix proteins. This finding suggests that hypoxia changes the microenvironment of the obese hypoxic adipose tissue by inducing the expression of not only adipokines but also peptidases such as CPA4 and CPE.

BolA Affects Cell Growth, and Binds to the Promoters of Penicillin-Binding Proteins 5 and 6 and Regulates Their Expression

  • Guinote, Ines Batista;Matos, Rute Goncalves;Freire, Patrick;Arraiano, Cecilia Maria
    • Journal of Microbiology and Biotechnology
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    • 제21권3호
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    • pp.243-251
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    • 2011
  • The gene bolA was discovered in the 80's, but unraveling its function in the cell has proven to be a complex task. The BolA protein has pleiotropic effects over cell physiology, altering growth and morphology, inducing biofilm formation, and regulating the balance of several membrane proteins. Recently, BolA was shown to be a transcription factor by repressing the expression of the mreB gene. The present report shows that BolA is a transcriptional regulator of the dacA and dacC genes, thus regulating both DD-carboxypeptidases PBP5 and PBP6 and thereby demonstrating the versatility of BolA as a cellular regulator. In this work, we also demonstrate that reduction of cell growth and survival can be connected to the overexpression of the bolA gene in different E. coli backgrounds, particularly in the exponential growth phase. The most interesting finding is that overproduction of BolA affects bacterial growth differently depending on whether the cells were inoculated directly from a plate culture or from an overnight batch culture. This strengthens the idea that BolA can be engaged in the coordination of genes that adapt the cell physiology in order to enhance cell adaptation and survival under stress conditions.