Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris
Yıl: 2017 Cilt: 41 Sayı: 5 Sayfa Aralığı: 746 - 753 Metin Dili: İngilizce İndeks Tarihi: 29-07-2022
Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris
Öz: The yeast Pichia pastoris expression system was investigated for the production of human cathepsin C (CatC) recombinant protein. The full-length CatC cDNA, corresponding to amino acids 12 475, was synthesized from interleukin-2 (IL-2) stimulated human peripheral blood mononuclear cells and subcloned in the pGEM-T cloning vector. After confirming the DNA sequence of the insert, the gene was cloned into the pPICZαA expression vector under the control of the methanol-inducible alcohol oxidase (AOX1) promoter and transformed to P. pastoris X-33 cells. The expressed protein was secreted into the culture medium through the α-factor mating signal sequence of the expression vector. Analysis of the culture supernatant revealed that the recombinant human CatC was secreted as a 58-kDa molecule, indicating that human CatC was accumulated in the culture supernatant as proform composed of the residual propart, the activation peptide, and the heavy and light chains. Extracellular recombinant proCatC was further activated by cysteine endoprotease papain in vitro and its activity was confirmed by assays using a synthetic substrate.
Anahtar Kelime: Konular:
Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık
- Adkinson AM, Raptis SZ, Kelley DG, Pham CT (2002). Dipeptidyl peptidase I activates neutrophil-derived serine proteases and regulates the development of acute experimental arthritis. J Clin Invest 109: 363-371.
- Brömme D, Li Z, Barnes M, Mehler E (1999). Human cathepsin V functional expression, tissue distribution, electrostatic surface potential, enzymatic characterization, and chromosomal localization. Biochemistry-US 38: 2377-2385.
- Cereghino JL, Cregg JM (2000). Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol Rev 24: 45-66.
- Cigic B, Krizaj I, Kralj B, Turk V, Pain RH (1998). Stoichiometry and heterogeneity of pro-region chain in tetrameric human cathepsin C. Biochim Biophys Acta 1382: 143-150.
- Cigic B, Pain RH (1999). Location of the binding site for chloride ion activation of cathepsin C. Eur J Biochem 264: 944-951.
- Coffey JW, de Duve C (1968). Digestive activity of lysosomes: I. The digestion of proteins by extracts of rat liver lysosomes. J Biol Chem 243: 3255-3263.
- Cregg JM, Vedvick TS, Raschke WC (1993). Recent advances in the expression of foreign genes in Pichia pastoris. Biotechnology 8: 905-910.
- Dahl SW, Halkier T, Lauritzen C, Dolenc I, Pedersen J, Turk V, Turk B (2001). Human recombinant pro-dipeptidyl peptidase I (cathepsin C) can be activated by cathepsins L and S but not by autocatalytic processing. Biochemistry-US 40: 1671-1678.
- Demain AL, Vaishnav P (2009). Production of recombinant proteins by microbes and higher organisms. Biotechnol Adv 27: 297- 306.
- Demirov D, Sarafian V, Kremensky I, Ganev V (1999). Evidence for protein splicing in the endoplasmic reticulum-Golgi intermediate compartment. Biochim Biophys Acta 1448: 507- 511.
- Dolenc I, Turk B, Pungercic G, Ritonja A, Turk V (1995). Oligomeric structure and substrate induced inhibition of human cathepsin C. J Biol Chem 270: 21626-21631.
- Izumya N, Fruton JS (1956). Specificity of cathepsin C. J Biol Chem 218: 59-76.
- Kim S, Nadel JA (2004). Role of neutrophils in mucus hypersecretion in COPD and implications for therapy. Treatments in Respiratory Medicine 3: 147-159.
- Lauritzen C, Pedersen J, Madsen MT, Justesen J, Martensen PM, Dahl SW (1998). Active recombinant rat dipeptidyl aminopeptidase I (cathepsin C) produced using the baculovirus expression system. Protein Expres Purif 14: 434-442.
- Linnevers CJ, McGrath ME, Armstrong R, Mistry FR, Barnes MG, Klaus JL, Palmer JT, Katz BA, Brömme D (1997). Expression of human cathepsin K in Pichia pastoris and preliminary crystallographic studies of an inhibitor complex. Protein Sci 4: 919-921.
- Methot N, Guay D, Rubin J, Ethier D, Ortega K, Wong S, Normandin D, Beaulieu C, Reddy TJ, Reindeau D et al. (2008). In vivo inhibition of serine protease processing requires a high fractional inhibition of cathepsin C. Mol Pharmacol 73: 1857- 1865.
- Methot N, Rubin J, Guay D, Beaulieu C, Ethier D, Jagadeeswar RT, Reindeau D, Percival MD (2007). Inhibition of the activation of multiple serine proteases with a cathepsin C inhibitor requires sustained exposure to prevent proenzyme processing. J Biol Chem 282: 20836-20846.
- Muno D, Ishidoh K, Ueno T, Kominami E (1993). Processing and transport of the precursor of cathepsin C during its transfer into lysosomes. Arch Biochem Biophys 306: 103-110.
- Paris A, Strukelj B, Pungercar J, Renko M, Dolenc I, Turk V (1995). Molecular cloning and sequence analysis of human preprocathepsin C. FEBS Lett 369: 326-330.
- Pedersen J, Lauritzen C, Madsen MT, Weis DS (1999). Removal of N-terminal polyhistidine tags from recombinant proteins using engineered aminopeptidases. Protein Expres Purif 15: 389-400.
- Rao NV, Rao GV, Hoidal JR (1997). Human dipeptidyl-peptidase I. Gene characterization, localization, and expression. J Biol Chem 272: 10260-0265.
- Wang B, Shi GP, Yao PM, Li Z, Chapman HA, Brömme D (1998). Human cathepsin F. Molecular cloning, functional expression, tissue localization, and enzymatic characterization. J Biol Chem 273: 32000-32008.
- Wolters PJ, Laig-Webster M, Caughey GH (2000). Dipeptidyl peptidase I cleaves matrix-associated proteins and is expressed mainly by mast cells in normal dog airways. Am J Resp Cell Mol 22: 183-190.
- Yang W, Xia W, Mao J, Xu D, Chen J, Feng S, Wang J, Li H, Theisen CF, Petersen JM et al. (2011). High level expression, purification and activation of human dipeptidyl peptidase I from mammalian cells. Protein Expres Purif 76: 59-64.
| APA | daglioglu c (2017). Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris. , 746 - 753. |
| Chicago | daglioglu cenk Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris. (2017): 746 - 753. |
| MLA | daglioglu cenk Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris. , 2017, ss.746 - 753. |
| AMA | daglioglu c Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris. . 2017; 746 - 753. |
| Vancouver | daglioglu c Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris. . 2017; 746 - 753. |
| IEEE | daglioglu c "Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris." , ss.746 - 753, 2017. |
| ISNAD | daglioglu, cenk. "Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris". (2017), 746-753. |
| APA | daglioglu c (2017). Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris. Turkish Journal of Biology, 41(5), 746 - 753. |
| Chicago | daglioglu cenk Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris. Turkish Journal of Biology 41, no.5 (2017): 746 - 753. |
| MLA | daglioglu cenk Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris. Turkish Journal of Biology, vol.41, no.5, 2017, ss.746 - 753. |
| AMA | daglioglu c Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris. Turkish Journal of Biology. 2017; 41(5): 746 - 753. |
| Vancouver | daglioglu c Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris. Turkish Journal of Biology. 2017; 41(5): 746 - 753. |
| IEEE | daglioglu c "Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris." Turkish Journal of Biology, 41, ss.746 - 753, 2017. |
| ISNAD | daglioglu, cenk. "Cloning, expression, and activity analysis of human cathepsin C in the yeast Pichia pastoris". Turkish Journal of Biology 41/5 (2017), 746-753. |
