title: Amino acids in the N-terminal region regulate the photocycle of photoactive yellow protein.
abstract: The spectroscopic properties of photoactive yellow protein (PYP) partially digested by chymotrypsin were studied. Chymotrypsin yielded three major products that were yellow but distinguishable by SDS-PAGE. They were readily separated by DEAE-Sepharose column chromatography. Protein sequencing and mass spectrometry demonstrated that chymotrypsin cleaved the N-terminal 6, 15, or 23 amino acids (T6, T15, and T23). The blue-shifts of the absorption maxima and the increases in the apparent pK(a) of the chromophores relative to those of intact PYP were less than 4 nm and 0.2, respectively. The absorption spectra of the near-UV intermediates produced from T6, T15, and T23 were identical to that of intact PYP, but with lifetimes that were 140, 2,300, and 4,500 times longer, respectively. These observations suggest that the recovery of the dark state of PYP from the near-UV intermediate is accelerated by the N-terminal region, and that this region acts as a regulatory factor for the photocycle of PYP.
authors: Graduate School of Materials Science, Nara Institute of Science and Technology, Ikoma, Nara 630-0101, Japan.
authors: M
authors: M
authors: Harigai
authors: S
authors: S
authors: Yasuda
authors: Y
authors: Y
authors: Imamoto
authors: K
authors: K
authors: Yoshihara
authors: F
authors: F
authors: Tokunaga
authors: M
authors: M
authors: Kataoka
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000528:radiation effects; D002852:Chromatography, Ion Exchange; D002918:Chymotrypsin / Q000378:metabolism; D020607:Halorhodospira halophila / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism* / Q000528:radiation effects; D010777:Photochemistry; D020283:Photoreceptors, Microbial; D013056:Spectrophotometry, Ultraviolet; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D010446:Peptide Fragments; D020283:Photoreceptors, Microbial; C050162:photoactive yellow protein, Bacteria; D002918:Chymotrypsin