The primary structure of a halorhodopsin from Natronobacterium pharaonis. Structural, functional and evolutionary implications for bacterial rhodopsins and halorhodopsins.
We cloned and sequenced the gene coding for the polypeptide of a halorhodopsin in Natronobacterium pharaonis (named here pharaonis halorhodopsin). Peptide sequencing of cyanogen bromide fragments, and immunoreactions of the protein and synthetic peptides derived from the COOH-terminal gene sequence, confirmed that the open reading frame is the structural gene for the pharaonis halorhodopsin polypeptide. The flanking DNA sequences, as well as those for other bacterial rhodopsins, were compared to previously proposed archaebacterial consensus sequences. In pairwise comparisons of the open reading frame with DNA sequences for bacterio-opsin and halo-opsin from Halobacterium halobium, silent divergences (mutations/nucleotide at codon positions which do not result in amino acid changes) were calculated. These indicate very considerable evolutionary distance between each pair of genes. In spite of this, the three protein sequences show extensive similarities, indicating strong selective pressures. Conserved and conservatively replaced amino acid residues in all three proteins identify general features essential for ion-motive bacterial rhodopsins, responsible for overall structure and chromophore properties. Comparison of the bacteriorhodopsin sequence with those of the two halorhodopsins, on the other hand, identifies features involved in their specific (proton and chloride ion) transport functions.
Lanyi
J K
JK
Department of Physiology and Biophysics, University of California, Irvine 92717.
Duschl
A
A
Hatfield
G W
GW
May
K
K
Oesterhelt
D
D
D000595:Amino Acid Sequence; D001105:Archaea / Q000032:analysis*; D001419:Bacteria / Q000032:analysis*; D001436:Bacteriorhodopsins / Q000276:immunology; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D006216:Halobacteriaceae / Q000032:analysis; D025602:Halorhodopsins; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D012045:Regulatory Sequences, Nucleic Acid; D013329:Structure-Activity Relationship; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic
D004269:DNA, Bacterial; D025602:Halorhodopsins; D008565:Membrane Proteins; D010446:Peptide Fragments; D001436:Bacteriorhodopsins