PMID- 22442222
OWN - NLM
STAT- MEDLINE
DCOM- 20120511
LR  - 20211021
IS  - 1744-3091 (Electronic)
IS  - 1744-3091 (Linking)
VI  - 68
IP  - Pt 3
DP  - 2012 Mar 1
TI  - Crystallization and preliminary X-ray crystallographic analysis of a 
      blue-light-absorbing proteorhodopsin.
PG  - 281-3
LID - 10.1107/S1744309111043612 [doi]
AB  - Proteorhodopsins (PRs), seven-transmembrane chromoproteins with retinal as a 
      chromophore, are light-driven proton pumps. To elucidate the light-driven 
      proton-pumping mechanism of PRs, a pET28a vector containing the 
      blue-light-absorbing proteorhodopsin (BPR) gene was constructed and the protein 
      was overexpressed in Escherichia coli. The protein was purified by immobilized 
      metal-ion affinity chromatography (IMAC). The purified BPR D97N mutant protein 
      (BPR_D97N) was crystallized using the vapour-diffusion method. Preliminary X-ray 
      diffraction data analysis showed that the crystal belonged to the orthorhombic 
      space group P2(1)2(1)2, with unit-cell parameters a = 161.6, b = 168.6, c = 64.7 
      A. A complete data set was collected to 3.3 A resolution using synchrotron 
      radiation on beamline X06 of the Swiss Light Source (SLS). Molecular replacement 
      was unsuccessful. To solve the structure of BPR_D97N by experimental phasing, 
      selenomethionine-substituted protein crystals were prepared. These crystals 
      diffracted to 3.0 A resolution and a complete data set was collected on beamline 
      BL17U of the Shanghai Synchrotron Radiation Facility (SSRF). Heavy-atom 
      substructure determination and phasing by SAD clearly showed that the crystal 
      contained five molecules in the asymmetric unit, with a V(M) of 3.26 A(3) Da(-1) 
      and a solvent content of 62.3%.
CI  - (c) 2012 International Union of Crystallography
FAU - Wang, Ning
AU  - Wang N
AD  - Key Laboratory of Agricultural and Environmental Microbiology, Ministry of 
      Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing 
      210095, People's Republic of China.
FAU - Wang, Meitian
AU  - Wang M
FAU - Gao, Yanyan
AU  - Gao Y
FAU - Ran, Tingting
AU  - Ran T
FAU - Lan, Yanli
AU  - Lan Y
FAU - Wang, Jian
AU  - Wang J
FAU - Xu, Langlai
AU  - Xu L
FAU - Wang, Weiwu
AU  - Wang W
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120222
PL  - England
TA  - Acta Crystallogr Sect F Struct Biol Cryst Commun
JT  - Acta crystallographica. Section F, Structural biology and crystallization 
      communications
JID - 101226117
RN  - 0 (Rhodopsins, Microbial)
RN  - 0 (proteorhodopsin)
RN  - 9009-81-8 (Rhodopsin)
SB  - IM
MH  - Crystallization
MH  - Crystallography, X-Ray
MH  - Mutation
MH  - Rhodopsin/analysis/*chemistry/genetics
MH  - Rhodopsins, Microbial
PMC - PMC3310530
EDAT- 2012/03/24 06:00
MHDA- 2012/05/12 06:00
PMCR- 2014/03/01
CRDT- 2012/03/24 06:00
PHST- 2011/07/22 00:00 [received]
PHST- 2011/10/20 00:00 [accepted]
PHST- 2012/03/24 06:00 [entrez]
PHST- 2012/03/24 06:00 [pubmed]
PHST- 2012/05/12 06:00 [medline]
PHST- 2014/03/01 00:00 [pmc-release]
AID - S1744309111043612 [pii]
AID - dp5009 [pii]
AID - 10.1107/S1744309111043612 [doi]
PST - ppublish
SO  - Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012 Mar 1;68(Pt 3):281-3. doi: 
      10.1107/S1744309111043612. Epub 2012 Feb 22.