Low-temperature spectroscopy of Met100Ala mutant of photoactive yellow protein

Photochemistry and Photobiology
Yasushi ImamotoMikio Kataoka

Abstract

The trans-to-cis photoisomerization of the p-coumaroyl chromophore of photoactive yellow protein (PYP) triggers the photocycle. Met100, which is located in the vicinity of the chromophore, is a key residue for the cis-to-trans back-isomerization of the chromophore, which is a rate-determining reaction of the PYP photocycle. Here we characterized the photocycle of the Met100Ala mutant of PYP (M100A) by low temperature UV-visible spectroscopy. Irradiation of M100A at 80 K yielded a 380 nm species (M100A(BL)), while the corresponding intermediate of wild type (WT; PYP(BL)) is formed above 90 K. The amounts of redshifted intermediates produced from M100A (M100A(B') and M100A(L)) were substantially less than those from WT. While the near-UV intermediate (PYP(M)) is not formed from WT in glycerol samples at low temperature, M100A(M) was clearly observed above 190 K. These alterations of the photocycle of M100A were explained by the shift in the equilibrium between the intermediates. The carbonyl oxygen of the thioester linkage of the cis-chromophore in the photocycle intermediates is close to the phenyl ring of Phe96 (<3.5 A), which would be displaced by the mutation of Met100. These findings imply that the interaction between chromo...Continue Reading

References

May 1, 1997·Journal of Biochemistry·K MiharaF Tokunaga
Feb 6, 2002·Biochemistry·Jun SasakiFumio Tokunaga
Nov 13, 2002·Biochemistry·Yasushi ImamotoMikio Kataoka
Jul 23, 2003·Nature Structural Biology·Elizabeth D GetzoffUlrich K Genick
Mar 18, 2004·The Journal of Biological Chemistry·Remco KortRaimond B G Ravelli
May 5, 2005·Proceedings of the National Academy of Sciences of the United States of America·Hyotcherl IheeKeith Moffat
Aug 2, 2006·Photochemistry and Photobiology·Tomokazu MorishitaYasushi Imamoto
Aug 31, 2006·Photochemistry and Photobiology·Yasushi Imamoto, Mikio Kataoka

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Citations

Jul 24, 2015·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·A KumarG A Woolley
Oct 19, 2010·The Journal of Physical Chemistry Letters·Elizabeth C CarrollDelmar S Larsen

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