Fourier transform infrared spectroscopic evidence for the existence of two conformations of the bacteriorhodopsin primary photoproduct at low temperature.

Fourier transform infrared difference spectroscopy of bacteriorhodopsin at low temperature reveals at least two stable forms of bacteriorhodopsin570 and the K photoproduct. In the case of bacteriorhodopsin570, warming from 81 to 135 K causes a reduction in absorption of several chromophore vibrations, but not the C = N stretching mode. These changes are consistent with a reorientation of the chromophore which leaves the angle of the C = N bond unchanged relative to the membrane plane. In the case of the K intermediate, two different forms can be isolated at 135 K on the basis of wavelength-dependent photoalteration. One form is identical to the low temperature K630 species, whereas a second blue-shifted form is present only above 135 K. This new form exhibits a 985 cm-1 peak in the hydrogen-out-of-plane bending region, which is similar to a reported room-temperature resonance Raman spectrum of K. Temperature-dependent changes in the conformation of the protein involving possible alterations in peptide hydrogen bonding are also detected.