PRIMARY STRUCTURE OF A PHOTOACTIVE YELLOW PROTEIN FROM THE PHOTOTROPHIC BACTERIUM ECTOTHIORHODOSPIRA-HALOPHILA, WITH EVIDENCE FOR THE MASS AND THE BINDING-SITE OF THE CHROMOPHORE

被引:84
作者
VANBEEUMEN, JJ
DEVREESE, BV
VANBUN, SM
HOFF, WD
HELLINGWERF, KJ
MEYER, TE
MCREE, DE
CUSANOVICH, MA
机构
[1] UNIV AMSTERDAM, DEPT MICROBIOL, AMSTERDAM, NETHERLANDS
[2] UNIV AMSTERDAM, CTR BIOTECHNOL, AMSTERDAM, NETHERLANDS
[3] UNIV ARIZONA, DEPT BIOCHEM, TUCSON, AZ 85721 USA
[4] Scripps Res Inst, RES INST, DEPT MOLEC BIOL, LA JOLLA, CA 92037 USA
关键词
AMINO ACID SEQUENCE; CHROMOPHORE STRUCTURE; ELECTROSPRAY MASS ANALYSIS; PHOTOACTIVE YELLOW PROTEIN;
D O I
10.1002/pro.5560020706
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The complete amino acid sequence of the 125-residue photoactive yellow protein (PYP) from Ectothiorhodospira halophila has been determined to be MEHVAFGSEDIENTLAKMDDGQLDGLAFGAIQLDGDGNILQYNAAEGDITGRDPKEVIGKNFFKDVAP CTDSPEFYGKFKEGVASGNLNTMFEYTFDYQMTPTKVKVHMKKALSGDSYWVFVKRV. This is the first sequence to be reported for this class of proteins. There is no obvious sequence homology to any other protein, although the crystal structure, known at 2.4 angstrom resolution (McRee, D.E., et al., 1989, Proc. Natl. Acad. Sci. USA 86, 6533-6537), indicates a relationship to the similarly sized fatty acid binding protein (FABP), a representative of a family of eukaryotic proteins that bind hydrophobic molecules. The amino acid sequence exhibits no greater similarity between PYP and FABP than for proteins chosen at random (8%). The photoactive yellow protein contains an unidentified chromophore that is bleached by light but recovers within a second. Here we demonstrate that the chromophore is bound covalently to Cys 69 instead of Lys 111 as deduced from the crystal structure analysis. The partially exposed side chains of Tyr 76, 94, and 118, plus Trp 119 appear to be arranged in a cluster and probably become more exposed due to a conformational change of the protein resulting from light-induced chromophore bleaching. The charged residues are not uniformly distributed on the protein surface but are arranged in positive and negative clusters on opposite sides of the protein. The exact chemical nature of the chromophore remains undetermined, but we here propose a possible structure based on precise mass analysis of a chromophore-binding peptide by electrospray ionization mass spectrometry and on the fact that the chromophore can be cleaved off the apoprotein upon reduction with a thiol reagent. The molecular mass of the chromophore, including an SH group, is 147.6 Da (+/-0.5 Da); the cysteine residue to which it is bound is at sequence position 69.
引用
收藏
页码:1114 / 1125
页数:12
相关论文
共 31 条
  • [1] AN INTRIGUING MEMBER OF THE LIPOCALIN PROTEIN FAMILY - ALPHA-1-MICROGLOBULIN
    AKERSTROM, B
    LOGDBERG, L
    [J]. TRENDS IN BIOCHEMICAL SCIENCES, 1990, 15 (06) : 240 - 243
  • [2] CLONING OF A CDNA-ENCODING RAT INTESTINAL FATTY-ACID BINDING-PROTEIN
    ALPERS, DH
    STRAUSS, AW
    OCKNER, RK
    BASS, NM
    GORDON, JI
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1984, 81 (02): : 313 - 317
  • [3] PLASMA DESORPTION MASS-SPECTROMETRY - COMING OF AGE
    COTTER, RJ
    [J]. ANALYTICAL CHEMISTRY, 1988, 60 (13) : A781 - &
  • [4] ELECTROSPRAY IONIZATION-PRINCIPLES AND PRACTICE
    FENN, JB
    MANN, M
    MENG, CK
    WONG, SF
    WHITEHOUSE, CM
    [J]. MASS SPECTROMETRY REVIEWS, 1990, 9 (01) : 37 - 70
  • [5] GOTSCHLICH EC, 1987, FEMS MICROBIOL LETT, V43, P253
  • [6] MODEL FOR THE STRUCTURE OF BACTERIORHODOPSIN BASED ON HIGH-RESOLUTION ELECTRON CRYOMICROSCOPY
    HENDERSON, R
    BALDWIN, JM
    CESKA, TA
    ZEMLIN, F
    BECKMANN, E
    DOWNING, KH
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 1990, 213 (04) : 899 - 929
  • [7] LOW-TEMPERATURE ABSORBENCY AND FLUORESCENCE SPECTROSCOPY OF THE PHOTOACTIVE YELLOW PROTEIN FROM ECTOTHIORHODOSPIRA-HALOPHILA
    HOFF, WD
    KWA, SLS
    VANGRONDELLE, R
    HELLINGWERF, KJ
    [J]. PHOTOCHEMISTRY AND PHOTOBIOLOGY, 1992, 56 (04) : 529 - 539
  • [8] THE MOLECULAR-STRUCTURE OF INSECTICYANIN FROM THE TOBACCO HORNWORM MANDUCA-SEXTA L AT 2.6 A RESOLUTION
    HOLDEN, HM
    RYPNIEWSKI, WR
    LAW, JH
    RAYMENT, I
    [J]. EMBO JOURNAL, 1987, 6 (06) : 1565 - 1570
  • [9] ADIPOCYTE P2 GENE - DEVELOPMENTAL EXPRESSION AND HOMOLOGY OF 5'-FLANKING SEQUENCES AMONG FAT CELL-SPECIFIC GENES
    HUNT, CR
    RO, JHS
    DOBSON, DE
    MIN, HY
    SPIEGELMAN, BM
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (11) : 3786 - 3790
  • [10] FATTY ACYLATED PROTEINS AS COMPONENTS OF INTRACELLULAR SIGNALING PATHWAYS
    JAMES, G
    OLSON, EN
    [J]. BIOCHEMISTRY, 1990, 29 (11) : 2623 - 2634