Arginine activity in the proton-motive photocycle of bacteriorhodopsin: solid-state NMR studies of the wild-type and D85N proteins

Biochemistry. 1999 Feb 2;38(5):1562-72. doi: 10.1021/bi981968z.

Abstract

15N solid-state NMR (SSNMR) spectra of guanidyl-15N-labeled bacteriorhodopsin (bR) show perturbation of an arginine residue upon deprotonation of the retinal Schiff base during the photocycle. At the epsilon position, an upfield shift of 4 ppm is observed while the eta nitrogens develop a pair of 'wing' peaks separated by 24 ppm. Proton-driven spin diffusion between the two 'wing' peaks indicates that they arise from a single Arg residue. An unusually asymmetric environment for this residue is indicated by comparison with guanidyl-15N chemical shifts in a series of arginine model compounds. The 'wing' peaks are tentatively assigned to Arg-82 on the basis of the SSNMR investigations of the alkaline and neutral dark-adapted forms of the D85N bacteriorhodopsin mutant. Another, less asymmetric pair of eta signals, that is not affected by Schiff base deprotonation or D85 mutation, is tentatively assigned to Arg-134. The results are discussed in relation to existing models of bR structure and function.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Substitution / genetics
  • Arginine / chemistry
  • Arginine / metabolism*
  • Asparagine / genetics*
  • Aspartic Acid / genetics*
  • Bacteriorhodopsins / chemistry
  • Bacteriorhodopsins / genetics
  • Bacteriorhodopsins / metabolism*
  • Carbon Isotopes
  • Crystallography, X-Ray
  • Halobacterium salinarum
  • Magnetic Resonance Spectroscopy
  • Photochemistry
  • Proton-Motive Force*

Substances

  • Carbon Isotopes
  • Aspartic Acid
  • Bacteriorhodopsins
  • Asparagine
  • Arginine