Isolation of photosystem I particles from spinach.
A method to prepare photosystem I (PSI) particles is described. Spinach leaves are used to prepare broken chloroplasts that are then solubilized by using a detergent (Triton X-100). Solubilized chloroplasts are then applied on an ion-exchange column. Eluted by a linear concentration gradient of NaCl, fractions enriched in PSI particles are collected and applied on a small hydroxyapatite column. By eluting with phosphate buffer, a concentrated preparation of PSI particles is obtained. The particles consist of PsaA, PsaB, PsaC, PsaD, PsaE, and PsaY. Assay methods that involve SDS-PAGE and P700 determination are also presented.
Hiyama
Tetsuo
T
Department of Biochemistry and Molecular Biology, Saitama University, Japan.
D002736:Chloroplasts / Q000737:chemistry; D002845:Chromatography / Q000379:methods; D004586:Electrophoresis / Q000379:methods; D045331:Photosystem I Protein Complex / Q000302:isolation & purification*; D010940:Plant Proteins / Q000032:analysis / Q000302:isolation & purification; D018724:Spinacia oleracea / Q000737:chemistry*
D045331:Photosystem I Protein Complex; D010940:Plant Proteins