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Characteristics of a de novo designed protein
Author(s) -
Tanaka Toshiki,
Kimura Hiromi,
Hayashi Mayumi,
Fujiyoshi Yosinori,
Fukuhara KenIchi,
Nakamura Haruki
Publication year - 1994
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560030306
Subject(s) - computational biology , biology
A series of 204 amino acid proteins intended to form TIM (triose phosphate isomerase) barrel structures were designed de novo. Each protein was synthesized by expression of the synthetic gene as a fusion protein with a portion of human growth hormone in an Escherichia coli host. After BrCN treatment, the protein was purified to homogeneity. The refolded proteins are globular and exist as monomers. One of the designed proteins is stable toward guanidine hydrochloride (GuHCl) denaturation, with a midpoint of 2.6 M determined from CD and tryp‐tophan fluorescence measurements. The GuHCl denaturation is well described by a 2‐state model. The NMR spectra, the thermal denaturation curves, and the 1‐anilino‐8‐naphthalene sulfonic acid binding imply that the stability of the protein arises mainly from hydrophobic interactions, which are probably of a nonspecific nature. The protein has a similar shape to that of rabbit triosephosphate isomerase, as determined by electron microscopy.