Development and evaluation of data-driven designed tags (DDTs) for controlling protein solubility.
Production of proteins is an important issue in protein science and pharmaceutical studies. Numerous protein expression systems using living cells and cell-free methods have been developed to date. In these systems, a promising strategy for improving the success rate of obtaining soluble proteins is the attachment of various tags into target proteins based on empirical rules. This paper presents a method for the production of data-driven designed tags (DDTs) based on highly frequent sequence property patterns in an experimentally assessed protein solubility dataset in a wheat germ cell-free system. We constructed seven proteins combined with 12 kinds of DDTs (six for enhancing solubility and six for insolubility) at the N-terminal region as tags. Then we investigated their behavior using SDS-PAGE. Results show that three and four proteins respectively showed a trend toward solubilization and insolubilization, which indicates the possibility that the theoretically designed sequence can control protein solubility.
Hirose
Shuichi
S
Computational Biology Research Center (CBRC), National Institute of Advanced Industrial Science and Technology (AIST), Tokyo, Japan. hirose-shuichi@aist.go.jp
Kawamura
Yoshifumi
Y
Mori
Masatoshi
M
Yokota
Kiyonobu
K
Noguchi
Tamotsu
T
Goshima
Naoki
N
D000595:Amino Acid Sequence; D002474:Cell-Free System; D030562:Databases, Protein; D006801:Humans; D008969:Molecular Sequence Data; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012995:Solubility
D011506:Proteins