Structural characterization of the α-mating factor prepro-peptide for secretion of recombinant proteins in Pichia pastoris.

Autor: Chahal S; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Wei P; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Moua P; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Park SP; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Kwon J; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Patel A; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Vu AT; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Catolico JA; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Tsai YF; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Shaheen N; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Chu TT; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Tam V; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Khan ZE; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Joo HH; Department of Chemistry, University of the Pacific, Stockton, CA 95211, USA., Xue L; Department of Chemistry, University of the Pacific, Stockton, CA 95211, USA., Lin-Cereghino J; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA., Tsai JW; Department of Chemistry, University of the Pacific, Stockton, CA 95211, USA., Lin-Cereghino GP; Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA. Electronic address: glincere@pacific.edu.
Jazyk: angličtina
Zdroj: Gene [Gene] 2017 Jan 20; Vol. 598, pp. 50-62. Date of Electronic Publication: 2016 Oct 29.
DOI: 10.1016/j.gene.2016.10.040
Abstrakt: The methylotrophic yeast Pichia pastoris has been used extensively for expressing recombinant proteins because it combines the ease of genetic manipulation, the ability to provide complex posttranslational modifications and the capacity for efficient protein secretion. The most successful and commonly used secretion signal leader in Pichia pastoris has been the alpha mating factor (MATα) prepro secretion signal. However, limitations exist as some proteins cannot be secreted efficiently, leading to strategies to enhance secretion efficiency by modifying the secretion signal leader. Based on a Jpred secondary structure prediction and knob-socket modeling of tertiary structure, numerous deletions and duplications of the MATα prepro leader were engineered to evaluate the correlation between predicted secondary structure and the secretion level of the reporters horseradish peroxidase (HRP) and Candida antarctica lipase B. In addition, circular dichroism analyses were completed for the wild type and several mutant pro-peptides to evaluate actual differences in secondary structure. The results lead to a new model of MATα pro-peptide signal leader, which suggests that the N and C-termini of MATα pro-peptide need to be presented in a specific orientation for proper interaction with the cellular secretion machinery and for efficient protein secretion.
(Copyright © 2016 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: