| Title: | Structural characterization of the alpha-mating factor prepro-peptide for secretion of recombinant proteins in Pichia pastoris |
| Author(s): | Chahal S; Wei P; Moua P; Park SP; Kwon J; Patel A; Vu AT; Catolico JA; Tsai YF; Shaheen N; Chu TT; Tam V; Khan ZE; Joo HH; Xue L; Lin-Cereghino J; Tsai JW; Lin-Cereghino GP; |
| Address: | "Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA. Department of Chemistry, University of the Pacific, Stockton, CA 95211, USA. Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA. Electronic address: glincere@pacific.edu" |
| DOI: | 10.1016/j.gene.2016.10.040 |
| ISSN/ISBN: | 1879-0038 (Electronic) 0378-1119 (Linking) |
| 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 (MATalpha) 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 MATalpha 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 MATalpha pro-peptide signal leader, which suggests that the N and C-termini of MATalpha pro-peptide need to be presented in a specific orientation for proper interaction with the cellular secretion machinery and for efficient protein secretion" |
| Keywords: | "Amino Acid Sequence Circular Dichroism Electrophoresis, Polyacrylamide Gel Fungal Proteins/chemistry/*genetics/metabolism Horseradish Peroxidase/genetics/metabolism Lipase/genetics/metabolism Mating Factor/chemistry/*genetics/metabolism Models, Molecular;" |
| Notes: | "MedlineChahal, Sabreen Wei, Peter Moua, Pachai Park, Sung Pil James Kwon, Janet Patel, Arth Vu, Anthony T Catolico, Jason A Tsai, Yu Fang Tina Shaheen, Nadia Chu, Tiffany T Tam, Vivian Khan, Zill-E-Huma Joo, Hyun Henry Xue, Liang Lin-Cereghino, Joan Tsai, Jerry W Lin-Cereghino, Geoff P eng Netherlands 2016/12/17 Gene. 2017 Jan 20; 598:50-62. doi: 10.1016/j.gene.2016.10.040. Epub 2016 Oct 29" |
