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

Authors

Sabreen Chahal, University of the Pacific
Peter Wei, University of the Pacific
Pachai Moua, University of the Pacific
Sung Park, University of the Pacific
Janet Kwon, University of the Pacific
Arth Patel, University of the Pacific
Anthony Vu, University of the Pacific
Jason Catolico, University of the Pacific
Yu Tsai, University of the Pacific
Nadia Shaheen, University of the Pacific
Tiffany Chu, University of the Pacific
Vivian Tam, University of the Pacific
Zill-E-Huma Khan, University of the Pacific
Hyun Joo, University of the Pacific
Liang Xue, University of the Pacific
Joan Lin-Cereghino, University of the PacificFollow
Jerry Tsai, University of the Pacific
Geoff P. Lin-Cereghino, University of the PacificFollow

Document Type

Article

Publication Title

Gene

Department

Biological Sciences

ISSN

0378-1119

Volume

598

DOI

10.1016/j.gene.2016.10.040

First Page

50

Last Page

62

Publication Date

1-20-2017

Abstract

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.

Recommended Citation

Chahal, S., Wei, P., Moua, P., Park, S., Kwon, J., Patel, A., Vu, A., Catolico, J., Tsai, Y., Shaheen, N., Chu, T., Tam, V., Khan, Z., Joo, H., Xue, L., Lin-Cereghino, J., Tsai, J., & Lin-Cereghino, G. P. (2017). Structural characterization of the α-mating factor prepro-peptide for secretion of recombinant proteins in Pichia pastoris. Gene, 598, 50–62. DOI: 10.1016/j.gene.2016.10.040
https://scholarlycommons.pacific.edu/cop-facarticles/718