Παρασκευή 25 Νοεμβρίου 2016

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

1-s2.0-S0378111916X00406-cov150h.gif

Publication date: 20 January 2017
Source:Gene, Volume 598
Author(s): Sabreen Chahal, Peter Wei, Pachai Moua, Sung Pil James Park, Janet Kwon, Arth Patel, Anthony T. Vu, Jason A. Catolico, Yu Fang Tina Tsai, Nadia Shaheen, Tiffany T. Chu, Vivian Tam, Zill-E-Huma Khan, Hyun Henry Joo, Liang Xue, Joan Lin-Cereghino, Jerry W. Tsai, Geoff P. Lin-Cereghino
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.



from #AlexandrosSfakianakis via Alexandros G.Sfakianakis on Inoreader http://ift.tt/2gsFmX3
via IFTTT

Δεν υπάρχουν σχόλια:

Δημοσίευση σχολίου

Δημοφιλείς αναρτήσεις