title: Deuterium solid-state nuclear magnetic resonance studies of methyl group dynamics in bacteriorhodopsin and retinal model compounds: evidence for a 6-s-trans chromophore in the protein.
abstract: Solid-state deuterium NMR spectroscopy is used to examine the dynamic behavior of 18-CD3 methyl groups in microcrystalline 6-s-cis-retinoic acid (triclinic) and 6-s-trans-retinoic acid (monoclinic) model compounds, as well as in the membrane protein bacteriorhodopsin (bR), regenerated with CD3-labeled retinal. Temperature dependent quadrupolar echo line shapes and T1 anisotropy measurements were used to characterize activation energies for 3-fold hopping motion of the methyl groups. These data provide supporting evidence that the conformation of the retinal chromophore in bR is 6-s-trans. The 6-s-cis conformer is characterized by strong eclipsing interactions between the 8-C proton and the 18-C methyl group protons; the 18-CD3 group shows an activation energy barrier for methyl 3-fold hopping of 14.5 +/- 1 kJ/mol. In contrast, the 18-CD3 group in the 6-s-trans isomer shows a considerably lower activation energy barrier of 5 +/- 1 kJ/mol. In bR, it is possible to obtain an approximate activation energy of 9 kJ/mol. This data is inconsistent with a 6-s-cis conformer but is consistent with the existence of a 6-s-trans-retinal Schiff base in bR with some interaction with the protein matrix. These results suggest that methyl rotor motions can be used to probe the van der Waals contact between a ligand and a protein binding pocket. The 6-s-trans conformer of the [16,17-(CD3)2]retinal in frozen hexane exhibits a major kinetic component with an activation energy barrier of of 14 -/+ 2 kJ/mol.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Francis Bitter National Magnet Laboratory, Massachusetts Institute of Technology, Cambridge 02139.
authors.forename: V
authors.initials: V
authors.lastname: Copié
authors.forename: A E
authors.initials: AE
authors.lastname: McDermott
authors.forename: K
authors.initials: K
authors.lastname: Beshah
authors.forename: J C
authors.initials: JC
authors.lastname: Williams
authors.forename: M
authors.initials: M
authors.lastname: Spijker-Assink
authors.forename: R
authors.initials: R
authors.lastname: Gebhard
authors.forename: J
authors.initials: J
authors.lastname: Lugtenburg
authors.forename: J
authors.initials: J
authors.lastname: Herzfeld
authors.forename: R G
authors.initials: RG
authors.lastname: Griffin
mesh_terms: D001436:Bacteriorhodopsins / Q000737:chemistry*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003198:Computer Simulation; D003460:Crystallization; D003903:Deuterium; D005615:Freezing; D006586:Hexanes; D009682:Magnetic Resonance Spectroscopy; D011487:Protein Conformation; D012172:Retinaldehyde / Q000737:chemistry*; D013696:Temperature; D013816:Thermodynamics; D014212:Tretinoin / Q000737:chemistry
chemical_list: D006586:Hexanes; D001436:Bacteriorhodopsins; D014212:Tretinoin; D003903:Deuterium; D012172:Retinaldehyde