2D-isolation of pure plasma and thylakoid membranes from the cyanobacterium Synechocystis sp. PCC 6803.
Aqueous polymer two-phase partitioning in combination with sucrose density centrifugation offered, for the first time, a 2D-separation method for the isolation of pure plasma and thylakoid membranes from the cyanobacterium Synechocystis 6803 without any cross-contaminations. The purity of the membrane fractions was verified by immunoblot analysis using antibodies against membrane-specific marker proteins. As an initiation of a proteomics project, two prominent proteins, which were observed only in the plasma membrane (Slr1513, a hypothetical protein, and HofG, a general secretion pathway protein), or in the thylakoid membrane (PsaE, a photosystem I protein, and NdhH, a subunit of NADH dehydrogenase), were identified.
Department of Biochemistry, Arrhenius Laboratories for Natural Sciences, Stockholm University, Sweden. birgitta@biokemi.su.se
B
B
Norling
E
E
Zak
B
B
Andersson
H
H
Pakrasi
D001426:Bacterial Proteins / Q000032:analysis / Q000737:chemistry; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000648:ultrastructure*; D002499:Centrifugation, Density Gradient / Q000379:methods; D000458:Cyanobacteria / Q000648:ultrastructure*; D003911:Dextrans; D007425:Intracellular Membranes / Q000648:ultrastructure*; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry; D015388:Organelles / Q000648:ultrastructure; D011092:Polyethylene Glycols; D012996:Solutions; D014461:Ultracentrifugation
D001426:Bacterial Proteins; D003911:Dextrans; D008565:Membrane Proteins; D012996:Solutions; D011092:Polyethylene Glycols