De novo design and creation of a stable artificial protein.
Protein de novo design has been performed, as an exercise of the inverse folding problem. A beta/alpha-barrel protein was designed and synthesized using the Escherichia coli expression system for the structural characterization. A tertiary model with a two-fold symmetry was built, based upon the geometrical parameters extracted from X-ray crystal structures of several beta/alpha-barrel proteins. Amino acid frequencies at each position on the alpha- and beta-structures were investigated, and an amino acid sequence with 201 residues was designed. The associated gene was chemically synthesized and the fusion protein with human growth hormone was expressed in Escherichia coli. The purified protein after being cleaved and refolded was found to be stable and globular with the large amount of secondary structures. However, it has similar characteristics to the molten globules of natural proteins, with loose packing of side-chains. The approach for the tight packing is discussed.
Tanaka
T
T
Protein Engineering Research Institute, Osaka, Japan.
Hayashi
M
M
Kimura
H
H
Oobatake
M
M
Nakamura
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001230:Aspergillus; D001426:Bacterial Proteins / Q000737:chemistry; D001483:Base Sequence; D002645:Chickens; D004355:Drug Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry; D008956:Models, Chemical; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry* / Q000235:genetics; D012475:Salmonella; D014305:Triose-Phosphate Isomerase / Q000737:chemistry; D014367:Tryptophan Synthase / Q000737:chemistry; D000516:alpha-Amylases / Q000737:chemistry
D001426:Bacterial Proteins; D005656:Fungal Proteins; D011506:Proteins; D000516:alpha-Amylases; D014367:Tryptophan Synthase; D014305:Triose-Phosphate Isomerase