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Influences of amino acid features of glutathione S ‐transferase fusion proteins on their solubility
Author(s) -
Shimada Kiyo,
Nagano Mihoko,
Kawai Makoto,
Koga Hisashi
Publication year - 2005
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200402085
Subject(s) - solubility , glutathione s transferase , chemistry , glutathione , fusion protein , biochemistry , transferase , fusion , enzyme , organic chemistry , recombinant dna , philosophy , gene , linguistics
We have previously described our strategy for high‐throughput (HT) production of recombinant antigens for anti‐mKIAA antibody generation, which involves using shotgun fragments generated during entire sequencing of mKIAA cDNAs. We applied this strategy to 1628 mouse KIAA (mKIAA) cDNA fragments, and 84.2% of the GST‐mKIAA fusion proteins were successfully purified. The solubility of the proteins was predicted by a small‐scale bacterial culture, and a large‐scale culture was then performed according to the expected results. Among them, 43.8% of the proteins were purified as a soluble form and 56.2% as an insoluble form. The average yield of the soluble proteins was 0.15 nmol/mL of bacterial culture, and that of the insoluble proteins was 0.55 nmol/mL Statistical analysis of the data revealed a significant correlation between amino acid features of the recombinant proteins and their solubility. To achieve the most effective and feasible protein expression, we constructed a decision tree in which the analyzed data were reflected. The information described here may provide practical guidelines for HT production of recombinant proteins.