Determination of the B820 subunit size of a bacterial core light-harvesting complex by small-angle neutron scattering.
The B820 subunit is an integral pigment-membrane protein complex and can be obtained by both dissociation of the core light-harvesting complex (LH1) in photosynthetic bacteria and reconstitution from its component parts in the presence of n-octyl beta-D-glucopyranoside (OG). Intrinsic size of the B820 subunit from Rhodospirillum rubrum LH1 complex was measured by small-angle neutron scattering in perdeuterated OG solution and evaluated by Guinier analysis. Both the B820 subunits prepared by dissociation of LH1 and reconstitution from apopolypeptides and pigments were shown to have a molecular weight of 11,400 +/- 500 and radius of gyration of 11.0 +/- 1.0 A, corresponding to a heterodimer consisting of one pair of alphabeta-polypeptides and two bacteriochlorophyll a molecules. Molecular weights of micelles formed by OG alone in solutions were determined in a range from 30,000 to 50,000 over concentrations of 1-5% (w/v), and thus are much larger than that of the B820 subunit. Similar measurement on the pigment-depleted apopolypeptides revealed highly heterogeneous behavior in the OG solutions, indicating that aggregates with various sizes were formed. The result provides evidence that bacteriochlorophyll a molecules play a crucial role in stabilizing and maintaining the B820 subunits in the dimeric state in solution. Further measurements on individual alpha- and beta-polypeptides exhibited a marked difference in aggregation property between the two polypeptides. The alpha-polypeptides appear to be uniformly dissolved in OG solution in a monomeric form, whereas the beta-polypeptides favor a self-associated form and tend to form large aggregates even in the presence of detergent. The difference in aggregation tendency was discussed in relation to the different behavior between alpha- and beta-polypeptides in reconstitution with bacteriochlorophyll a molecules.
D001059:Apoproteins / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry; D001429:Bacteriochlorophylls / Q000737:chemistry; D002338:Carotenoids / Q000737:chemistry; D003903:Deuterium; D019281:Dimerization; D005960:Glucosides / Q000737:chemistry; D045342:Light-Harvesting Protein Complexes; D008823:Micelles; D008970:Molecular Weight; D033363:Neutron Diffraction / Q000379:methods; D009502:Neutrons; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D021122:Protein Subunits; D012247:Rhodospirillum rubrum / Q000737:chemistry*; D012542:Scattering, Radiation; D013053:Spectrophotometry / Q000379:methods
D001059:Apoproteins; D001426:Bacterial Proteins; D001429:Bacteriochlorophylls; D005960:Glucosides; D045342:Light-Harvesting Protein Complexes; D008823:Micelles; D045322:Photosynthetic Reaction Center Complex Proteins; D021122:Protein Subunits; C018619:octyl-beta-D-glucoside; D002338:Carotenoids; D003903:Deuterium