Crystallization and characterization of two crystal forms of the B800–850 light-harvesting complex from Rhodopseudomonas acidophila strain 10050

Papiz, Miroslav Z.; Hawthornthwaite, A. M.; Cogdell, Richard J.; Woolley, Kevin J.; Wightman, P.A.; Ferguson, Linda A.; Lindsay, J. Gordon

doi:10.1016/0022-2836(89)90612-8

Cited by 80 publications

(33 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Given the monomer molecular weight of 30 kDa, a packing with one molecule per asymmetric unit is most likely since this corresponds to a Matthews coefficient (VM) [21] of 3.2 A3/Da and 61% solvent content. This agrees well with values obtained for other membrane proteins [8,9,15 19], ranging from a V M of 2.5 ~3/Da (51% solvent content) for crystals of a light-harvesting complex [19] to a V M of 5.2/~3/Da (76% solvent content) for crystals of maltoporin from E. coli [16].…”

Section: Resultssupporting

confidence: 90%

See 1 more Smart Citation

Crystallization and preliminary X‐ray analysis of outer membrane phospholipase A fromEscherichia coli

et al. 1995

View full text Add to dashboard Cite

Section: Resultssupporting

confidence: 90%

“…The conditions used so far to crystallize membrane proteins employed either PEG or high salt concentrations as crystallizing agents [8,9,[14][15][16][17][18][19][20]. OMPLA from E. coli is the first membrane protein crystallized using an organic solvent such as MPD as crystallizing agent.…”

Section: Resultsmentioning

confidence: 99%

Crystallization and preliminary X‐ray analysis of outer membrane phospholipase A fromEscherichia coli

et al. 1995

View full text Add to dashboard Cite

“…Moderate amounts of organic amphiphiles have been shown to be advantageous for membrane protein crystallisation, and their beneficial effect was attributed to increased micellar fluidity and deformability and decreased micellar size (Garavito et al, 1986;Marone et al, 1999;Michel, 1982Michel, , 1983Papiz et al, 1989;Timmins et al, 1991). In higher concentrations (typical 20-25%) small amphiphiles even can induce a phase transition (Becher and Trifiletti, 1973).…”

Section: Crystal Packing and Crystal Growthmentioning

confidence: 99%

Detergent organisation in crystals of monomeric outer membrane phospholipase A

Snijder¹,

Timmins²,

Kalk³

et al. 2003

Journal of Structural Biology

View full text Add to dashboard Cite

The structure of the detergent in crystals of outer membrane phospholipase A (OMPLA) has been determined using neutron diffraction contrast variation. Large crystals were soaked in stabilising solutions, each containing a different H 2 O=D 2 O contrast. From the neutron diffraction at five contrasts, the 12 A A resolution structure of the detergent micelle around the protein molecule was determined. The hydrophobic b-barrel surfaces of the protein molecules are covered by rings of detergent. These detergent belts are fused to neighbouring detergent rings forming a continuous three-dimensional network throughout the crystal. The thickness of the detergent layer around the protein varies from 7-20 A A. The enzymeÕs active site is positioned just outside the hydrophobic detergent zone and is thus in a proper location to catalyse the hydrolysis of phospholipids in a natural membrane. Although the dimerisation face of OMPLA is covered with detergent, the detergent density is weak near the exposed polar patch, suggesting that burying this patch in the enzymeÕs dimer interface may be energetically favourable. Furthermore, these results indicate a crucial role for detergent coalescence during crystal formation and contribute to the understanding of membrane protein crystallisation.

show abstract

“…sphaeroides is known to atomic resolution (3,4); however, the structures of the LH1 and LH2 complexes are not known, although diffracting crystals ofvarious LH complexes have been reported (5)(6)(7)(8)(9)(10). As yet, these structures have not been solved, so it is necessary to employ other methods such as mutagenesis and spectroscopy to obtain relevant data.…”

mentioning

confidence: 99%

Modification of a hydrogen bond to a bacteriochlorophyll a molecule in the light-harvesting 1 antenna of Rhodobacter sphaeroides.

Olsen

Sockalingum

Robert

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

119

141

View full text Add to dashboard Cite

Site-directed mutagenesis has been used to examine the function of a highly conserved aromatic residue, darp43, in the light-harvesting 1 antenna ofthe photosynthetic bacterium Rhodobacter sphaeroides. In this antenna dfrp43 is thought to be located near the putative binding site for bacteriochlorophyll; in this work it was changed to both Tyr and The photosynthetic apparatus of the purple bacterium Rhodobacter sphaeroides consists of a membrane protein complex, the reaction center (RC), surrounded by a core light-harvesting complex, LH1, also referred to as B875, in a fixed stoichiometry of -12 LH1 a(3 dimers per RC. An additional peripheral light-harvesting complex, LH2, also referred to as B800-850, occurs in variable amounts in response to environmental levels of light and oxygen (1, 2). The structure of the RC of Rb. sphaeroides is known to atomic resolution (3, 4); however, the structures of the LH1 and LH2 complexes are not known, although diffracting crystals ofvarious LH complexes have been reported (5-10). As yet, these structures have not been solved, so it is necessary to employ other methods such as mutagenesis and spectroscopy to obtain relevant data. As a result of these approaches, more information is becoming available on the interaction of the chromophores of LH1 and LH2 with nearby residues of the antenna polypeptides (11,12). These data can be used to test existing model structures derived from spectroscopic and electrophoretic mobility studies of wild-type (WT) complexes. The models that have been proposed for bacterial LH complexes all follow a similar pattern in that the af8 heterodimer, which in the case of LH1 would bind 2 bacteriochlorophyll (bchl) molecules, is considered to be the minimum possible building block. The topology and conformation of these subunits are thought to be similar, each with a cytoplasmically exposed N terminus, a single transmembrane helix, and a periplasmic C terminus. There are indications of turns at or near the membrane interfaces (13 (20), and Rhodocyclus gelatinosus (21), leading to the speculation that this bchl dimer is the building block of the LH1 complex. Characterization of the B820 form (22) supports the idea that it is an excitonically coupled bchl a dimer. Moreover, resonance Raman measurements of the B820 showed that it differed from the B873 form in the interactions of the C2 acetyl carbonyl; the loss of a H bond was one possible explanation of the Raman data. A more detailed Raman study of the LH1 ofRs. rubrum G9 has shown that upon formation of the B820, the strength of one H bond was increased (23 FT, Fourier transform; WT, wild type; P-OG, n-octyl P-Dglucopyranoside; ODPS, n-octyldipropyl sulfoxide. 7124The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

show abstract

Crystallization and characterization of two crystal forms of the B800–850 light-harvesting complex from Rhodopseudomonas acidophila strain 10050

Cited by 80 publications

References 12 publications

Crystallization and preliminary X‐ray analysis of outer membrane phospholipase A fromEscherichia coli

Crystallization and preliminary X‐ray analysis of outer membrane phospholipase A fromEscherichia coli

Detergent organisation in crystals of monomeric outer membrane phospholipase A

Modification of a hydrogen bond to a bacteriochlorophyll a molecule in the light-harvesting 1 antenna of Rhodobacter sphaeroides.

Contact Info

Product

Resources

About