Structure-function studies of bacteriorhodopsin XV: Effects of deletions in loops B-C and E-F on bacteriorhodopsin chromophore and structure

Marie A. Gilles-Gonzalez, Donald M. Engelman, H. Gobind Khorana

Research output: Contribution to journalArticle

41 Scopus citations

Abstract

Bacteriorhodopsin mutants containing deletions in loop B-C, ΔThr67-Glu74 or ΔGly65-Gln75, or a deletion in the loop E-F, ΔGlu161-Ala168, were prepared. Following their expression in Escherichia coli, the mutant proteins were purified to homogeneity and refolded with retinal in detergent-phospholipid mixtures. The mutants containing deletions in the loop B-C were normal at 4 °C but showed the following changes at 20 °C. 1) The λ(max) shifted from 540 to below 510 nm; 2) the rates of bleaching by hydroxylamine in the dark increased; and 3) the rate and steady state of proton pumping decreased. Deletion of the eight amino acids in loop E-F did not affect wild-type behavior. However, all the mutant proteins were more prone to thermal and sodium dodecyl sulfate denaturation than the wild-type bacteriorhodopsin. These observations show that the structures of the B-C and E-F loops are not essential for correct folding of bacteriorhodopsin, but they contribute to the stability of the folded protein.

Original languageEnglish (US)
Pages (from-to)8545-8550
Number of pages6
JournalJournal of Biological Chemistry
Volume266
Issue number13
StatePublished - 1991

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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