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Journal of Protein Chemistry

, Volume 15, Issue 3, pp 281–289 | Cite as

Effect of transmembrane helix packing on tryptophan and tyrosine environments in detergent-solubilized bacterio-opsin

  • Robert Renthal
  • Patrick Haas
Article

Abstract

Bacterio-opsin (bO) is folded in a nearly native conformation in mixed micelles of dimyristoyl phosphatidyl choline (DMPC) and 3-[(3-cholamidopropyl)-dimehtylamonio]-1-propane sulfonic acid (CHAPS), but bO is partially unfolded in sodium dodecyl sulfate (SDS). UV difference spectroscopy was used to study the changes in environment of bO aromatic amino acid side chains that occur upon partial unfolding. The UV difference spectra of peptides in CHAPS/DMPC minus peptides in SDS were measured for bO and the following subfragments of bO: C1 (residues 72–248), C2 (1–71), V1 (1–166), V2 (167–248), CB7 (119–145), CB9 (164–209), and CB10 (72–118). The spectra show that, in partially unfolded bO in SDS, the Tyr and Trp absorbance is blue-shifted. The difference spectra were compared to solvent perturbation difference spectra of N-acetyl-l-tyrosine ethyl ester and N-acetyl-l-tryptophanamide. The exposure change calculated from the difference spectra was found to correlate with the change in the number of van der Waals contacting atoms upon partial unfolding, and also with the number of transmembrane helical segments. This result suggests a simple experimental method of testing helix packing arrangements derived from hydropathy plots and model building.

Key words

Bacteriorhodopsin protein folding UV difference spectroscopy detergent micelles proteolysis CNBr 

Abbreviations

bO

bacterio-opsin

bR

bacteriorhodopsin

SDS

sodium dodecyl sulfate

CHAPS

3-[(3-cholamidopropyl)-dimethylamonio]-1-propane sulfonic acid

C1

bacteriorhodopsin residues 72–248

C2

1-71

VI

1-166

V2

167-248

CB7

119-145

CB9

164-209

CB10

72-118

DMPC

1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine

EDTA

di-sodium ethylenediamine tetra-aceticacid

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Copyright information

© Plenum Publishing Corporation 1996

Authors and Affiliations

  1. 1.Division of Earth and Physical SciencesUniversity of Texas at San AntonioSan Antonio
  2. 2.Department of BiochemistryUniversity of Texas Health Science Center at San AntonioSan Antonio

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