The isolation and partial characterization of the pyrenoid protein of Eremosphaera viridis.
The pyrenoids of Eremosphaera viridis, a green alga, were isolated by density gradient centrifugation and their physical and enzymatic properties were studied. The ultraviolet absorption spectrum of sodium dodecyl sulfate (SDS) extracts of pyrenoids showed a single peak at a wavelength of 277 nm, indicating the presence of protein and the probable absence of nucleic acid. Upon electrophoresis on polyacrylamide gels containing SDS, 16 bands were resolved of which two, together, accounted for 90% of the total protein on the gels. The molecular weights of these two proteins were estimated to be 59,000 and 12,300 and the ratio by weight of the larger to the smaller protein was found to be 2:1. The physical and enzymatic properties of these two proteins were found to closely resemble the properties reported in the literature for the subunits of fraction I protein. Both pyrenoids and fraction I protein are localized in the chloroplast, and both have two principal protein components. The molecular weights and relative ratio of the two pyrenoid components are very similar to those of the two components of fraction I protein. The pyrenoid was found to contain a high specific activity of ribulose-1,5-diphosphate carboxylase which is the same enzymatic activity exhibited by fraction I protein. The presence of ribose-5-phosphate isomerase and ribulose-5-phosphate kinase activities was also noted in pyrenoid preparations. It is suggested that the pyrenoid contains fraction I protein and possibly other enzymes of the Calvin-Bassham carbon dioxide fixing pathway.
D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D000460:Chlorophyta / Q000166:cytology*; D002736:Chloroplasts / Q000032:analysis* / Q000201:enzymology; D003470:Culture Media; D004589:Electrophoresis, Disc; D005856:Germ-Free Life; D006651:Histocytochemistry; D007535:Isomerases / Q000032:analysis; D008025:Ligases / Q000032:analysis; D008854:Microscopy, Electron; D008970:Molecular Weight; D009940:Organoids / Q000032:analysis / Q000201:enzymology; D010770:Phosphotransferases / Q000032:analysis; D010940:Plant Proteins / Q000032:analysis / Q000302:isolation & purification*; D013057:Spectrum Analysis; D013213:Starch / Q000032:analysis; D014466:Ultraviolet Rays
D003470:Culture Media; D010940:Plant Proteins; D013213:Starch; D010770:Phosphotransferases; D007535:Isomerases; D008025:Ligases