Translated products of tandem microgene repeats exhibit diverse properties also seen in natural proteins.
Repetitiousness is often observed in the primary and tertiary structures of proteins. We are intrigued by the potential role played by periodicity in the evolution of proteins and have created artificial repetitious proteins from repeats of short DNA sequences (microgenes). In this paper we characterize the physicochemical properties of six such artificially created proteins, which are the translated products of repeats of three microgenes. Three of the six proteins contain beta-sheet-like structures and are rather hydrophobic in nature. These proteins form macroscopic membranous structures in the presence of monovalent cationic ions, suggesting they have the capacity to promote strong intermolecular interactions. Of the other three proteins, one is comprised of alpha-helices and two have disordered structures. Small angle X-ray scattering analysis indicates that the artificial proteins do not fold as tightly as natural proteins, but are more compact than if completely denatured. One alpha-helical protein whose microgene unit was designed from coiled coil proteins was crystallized, demonstrating that repetitious artificial proteins can undergo transition to a more ordered state under appropriate conditions. Application of this approach to the development of a novel protein engineering system is discussed.
Shiba
Kiyotaka
K
Department of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, Toshima, Tokyo 170-8455, Japan. kshiba@jfcr.or.jp
Shirai
Tsuyoshi
T
Honma
Takako
T
Noda
Tetsuo
T
D000595:Amino Acid Sequence; D003460:Crystallization; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000737:chemistry* / Q000235:genetics*; D010784:Photomicrography; D014176:Protein Biosynthesis / Q000235:genetics*; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D020080:Tandem Repeat Sequences / Q000235:genetics*
D010455:Peptides; D011994:Recombinant Proteins