Fourier transform infrared difference spectroscopy of rhodopsin mutants: light activation of rhodopsin causes hydrogen-bonding change in residue aspartic acid-83 during meta II formation. Export

@article{citeulike:4197395, abstract = {Fourier transform infrared (FTIR) difference spectroscopy and site-directed mutagenesis have been used to investigate structural changes which occur during rhodopsin photoactivation at the level of individual amino acid residues. The rhodopsin-->bathorhodopsin FTIR difference spectra of the mutants Asp-83-->Asn (D83N) and Glu-134-->Asp (E134D) incorporated into membranes are similar to that of native rhodopsin in the photoreceptor membrane, demonstrating that the retinal chromophores of these mutants undergo a normal 11-cis to all-trans photoisomerization. Two bands assigned to the C = O stretching mode of Asp and/or Glu carboxylic acid groups are absent in the D83N rhodopsin-->metarhodopsin II FTIR difference spectrum. Corresponding changes are not observed in the carboxylate C = O stretching region. The most straightforward explanation is that the carboxylic acid group of Asp-83 remains protonated in rhodopsin and its bleaching intermediates but undergoes an increase in its hydrogen bonding during the metarhodopsin I-->metarhodopsin II transition. The mutant E134D produced a normal rhodopsin-->bathorhodopsin and rhodopsin-->metarhodopsin II difference spectrum, but a fraction of misfolded protein was observed, supporting earlier evidence that Glu-134 plays a role in proper protein insertion and/or folding in the membrane.}, author = {Rath, P. and DeCaluw\'{e}, L. L. and Bovee-Geurts, P. H. and DeGrip, W. J. and Rothschild, K. J.}, citeulike-article-id = {4197395}, citeulike-linkout-0 = {http://view.ncbi.nlm.nih.gov/pubmed/8399169}, citeulike-linkout-1 = {http://www.hubmed.org/display.cgi?uids=8399169}, comment = {Evidence that D2.50 remains protonated in rhodopsin and its bleaching intermediates but undergoes an increase in its hydrogen bonding during the MI MII.}, day = {5}, issn = {0006-2960}, journal = {Biochemistry}, keywords = {activation, d250, protonation, rhodopsin, spectroscopy, switch}, month = {October}, number = {39}, pages = {10277--10282}, posted-at = {2009-03-20 11:55:44}, priority = {2}, title = {Fourier transform infrared difference spectroscopy of rhodopsin mutants: light activation of rhodopsin causes hydrogen-bonding change in residue aspartic acid-83 during meta II formation.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/8399169}, volume = {32}, year = {1993} }

TY - JOUR ID - citeulike:4197395 L3 - citeulike-article-id:4197395 N2 - Fourier transform infrared (FTIR) difference spectroscopy and site-directed mutagenesis have been used to investigate structural changes which occur during rhodopsin photoactivation at the level of individual amino acid residues. The rhodopsin-->bathorhodopsin FTIR difference spectra of the mutants Asp-83-->Asn (D83N) and Glu-134-->Asp (E134D) incorporated into membranes are similar to that of native rhodopsin in the photoreceptor membrane, demonstrating that the retinal chromophores of these mutants undergo a normal 11-cis to all-trans photoisomerization. Two bands assigned to the C = O stretching mode of Asp and/or Glu carboxylic acid groups are absent in the D83N rhodopsin-->metarhodopsin II FTIR difference spectrum. Corresponding changes are not observed in the carboxylate C = O stretching region. The most straightforward explanation is that the carboxylic acid group of Asp-83 remains protonated in rhodopsin and its bleaching intermediates but undergoes an increase in its hydrogen bonding during the metarhodopsin I-->metarhodopsin II transition. The mutant E134D produced a normal rhodopsin-->bathorhodopsin and rhodopsin-->metarhodopsin II difference spectrum, but a fraction of misfolded protein was observed, supporting earlier evidence that Glu-134 plays a role in proper protein insertion and/or folding in the membrane. IS - 39 JF - Biochemistry SN - 0006-2960 EP - 10282 TI - Fourier transform infrared difference spectroscopy of rhodopsin mutants: light activation of rhodopsin causes hydrogen-bonding change in residue aspartic acid-83 during meta II formation. VL - 32 SP - 10277 KW - activation KW - d250 KW - protonation KW - rhodopsin KW - spectroscopy KW - switch N1 - Evidence that D2.50 remains protonated in rhodopsin and its bleaching intermediates but undergoes an increase in its hydrogen bonding during the MI MII. AU - Rath, P AU - DeCaluwé, LL AU - Bovee-Geurts, PH AU - DeGrip, WJ AU - Rothschild, KJ PY - 1993/10/05/ UR - http://view.ncbi.nlm.nih.gov/pubmed/8399169 ER -