Crystallographic intermediates of structures of the M and N bacteriorhodopsin: Assembly of a hydrogen-bonded chain of water molecules between Asp-96 and the retinal Schiff base

被引:138
作者
Schobert, B [1 ]
Brown, LS [1 ]
Lanyi, JK [1 ]
机构
[1] Univ Calif Irvine, Dept Physiol & Biophys, Irvine, CA 92697 USA
关键词
bacteriorhodopsin; X-ray diffraction; hydrogen-bonded chain; M photointermediate; N photointermediate;
D O I
10.1016/S0022-2836(03)00576-X
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
An M intermediate of wild-type bacteriorhodopsin and an N intermediate of the V49A mutant were accumulated in photostationary states at pH 5.6 and 295 K, and their crystal structures determined to 1.52 Angstrom and 1.62 Angstrom resolution, respectively. They appear to be M-1 and N' in the sequence, M-1 <----> M-2 <----> M'(2) <----> N <----> N' --> O --> BR, where M-1, M-2, and M'(2) contain an unprotonated retinal Schiff base before and after a reorientation switch and after proton release to the extracellular surface, while N and M contain a reprotonated Schiff base, before and after reprotonation of Asp96 from the cytoplasmic surface. In M-1, we detect a cluster of three hydrogen-bonded water molecules at Asp96, not present in the BR state. In M-2, whose structure we reported earlier, one of these water molecules intercalates between Asp96 and Thr46. In M, the cluster is transformed into a single-file hydrogen-bonded chain of four water molecules that connects Asp96 to the Schiff base. We find a network of three water molecules near residue 219 in the crystal structure of the non-illuminated F219L mutant, where the residue replacement creates a cavity This suggests that the hydration of the cytoplasmic region we observe in M might have occurred spontaneously, beginning at an existing water molecule as nucleus, in the cavities from residue rearrangements in the photocycle. (C) 2003 Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:553 / 570
页数:18
相关论文
共 72 条
[51]   D38 is an essential part of the proton translocation pathway in bacteriorhodopsin [J].
Riesle, J ;
Oesterhelt, D ;
Dencher, NA ;
Heberle, J .
BIOCHEMISTRY, 1996, 35 (21) :6635-6643
[52]   Unraveling photoexcited conformational changes of bacteriorhodopsin by time resolved electron paramagnetic resonance spectroscopy [J].
Rink, T ;
Pfeiffer, M ;
Oesterhelt, D ;
Gerwert, K ;
Steinhoff, HJ .
BIOPHYSICAL JOURNAL, 2000, 78 (03) :1519-1530
[53]   Crystal structure of the D85S mutant of bacteriorhodopsin: Model of an O-like photocycle intermediate [J].
Rouhani, S ;
Cartailler, JP ;
Facciotti, MT ;
Walian, P ;
Needleman, R ;
Lanyi, JK ;
Glaeser, RM ;
Luecke, H .
JOURNAL OF MOLECULAR BIOLOGY, 2001, 313 (03) :615-628
[54]   Thermodynamic stability of water molecules in the bacteriorhodopsin proton channel: A molecular dynamics free energy perturbation study [J].
Roux, B ;
Nina, M ;
Pomes, R ;
Smith, JC .
BIOPHYSICAL JOURNAL, 1996, 71 (02) :670-681
[55]   Helix deformation is coupled to vectorial proton transport in the photocycle of bacteriorhodopsin [J].
Royant, A ;
Edman, K ;
Ursby, T ;
Pebay-Peyroula, E ;
Landau, EM ;
Neutze, R .
NATURE, 2000, 406 (6796) :645-648
[56]   Lipidic cubic phases: New matrices for the three-dimensional crystallization of membrane proteins [J].
Rummel, G ;
Hardmeyer, A ;
Widmer, C ;
Chiu, ML ;
Nollert, P ;
Locher, KP ;
Pedruzzi, I ;
Landau, EM ;
Rosenbusch, JP .
JOURNAL OF STRUCTURAL BIOLOGY, 1998, 121 (02) :82-91
[57]   Structural alterations for proton translocation in the M state of wild-type bacteriorhodopsin [J].
Sass, HJ ;
Büldt, G ;
Gessenich, R ;
Hehn, D ;
Neff, D ;
Schlesinger, R ;
Berendzen, J ;
Ormos, P .
NATURE, 2000, 406 (6796) :649-653
[58]   Subsecond proton-hole propagation in bacteriorhodopsin [J].
Schätzler, B ;
Dencher, NA ;
Tittor, J ;
Oesterhelt, D ;
Yaniv-Checover, S ;
Nachliel, E ;
Gutman, M .
BIOPHYSICAL JOURNAL, 2003, 84 (01) :671-686
[59]   Crystallographic structure of the K intermediate of bacteriorhodopsin: Conservation of free energy after photoisomerization of the retinal [J].
Schobert, B ;
Cupp-Vickery, J ;
Hornak, V ;
Smith, SO ;
Lanyi, JK .
JOURNAL OF MOLECULAR BIOLOGY, 2002, 321 (04) :715-726
[60]   SHELXL: High-resolution refinement [J].
Sheldrick, GM ;
Schneider, TR .
MACROMOLECULAR CRYSTALLOGRAPHY, PT B, 1997, 277 :319-343