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Origins of the Intermediate Spectral Form in M100 Mutants of Photoactive Yellow Protein
Author(s) -
Kumar Anil,
Woolley George Andrew
Publication year - 2015
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/php.12464
Subject(s) - chromophore , chemistry , hydrogen bond , absorbance , mutant , crystallography , photochemistry , stereochemistry , molecule , organic chemistry , biochemistry , gene , chromatography
Numerous single‐site mutants of photoactive yellow protein ( PYP ) from H alorhodospira halophila and as well as PYP homologs from other species exhibit a shoulder on the short wavelength side of the absorbance maximum in their dark‐adapted states. The structural basis for the occurrence of this shoulder, called the “intermediate spectral form,” has only been investigated in detail for the Y42F mutation. Here we explore the structural basis for occurrence of the intermediate spectral form in a M121E derivative of a circularly permuted H. halophila PYP (M121E‐ cPYP ). The M121 site in M121E‐ cPYP corresponds to the M100 site in wild‐type H. halophila PYP . High‐resolution NMR measurements with a salt‐tolerant cryoprobe enabled identification of those residues directly affected by increasing concentrations of ammonium chloride, a salt that greatly enhances the fraction of the intermediate spectra form. Residues in the surface loop containing the M121E (M100E) mutation were found to be affected by ammonium chloride as well as a discrete set of residues that link this surface loop to the buried hydroxyl group of the chromophore via a hydrogen bond network. Localized changes in the conformational dynamics of a surface loop can thereby produce structural rearrangements near the buried hydroxyl group chromophore while leaving the large majority of residues in the protein unaffected.