Functional proteomics of circadian expressed proteins from Chlamydomonas reinhardtii

[摘要]

In this study, functional proteomics was successfully applied for the characterization of circadian expressed, basic proteins. For this purpose, we have chosen the green model alga Chlamydomonas reinhardtii since its entire nuclear genome is available and it is ideally suited for biochemical enrichment procedures. Proteins from cells harvested during subjective day and night were heparin affinity purified. They were separated by two-dimensional gel electrophoresis suited for basic proteins and analyzed after tryptic digestion by electrospray ionization mass spectrometry. We can show for the first time that the expressions of a protein disulfide isomerase-like protein and a tetratricopeptide repeat protein change in a circadian manner. Interestingly, both proteins are known to be interaction partners in multiprotein complexes including RNA binding proteins.

[学科分类] 生物化学/生物物理

[关键词] Circadian rhythm;Functional proteomics;Protein disulfide isomerase;Tetratricopeptide repeat protein;Chlamydomonas reinhardtii;CHAPS;3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate;DTT;dithiothreitol;ESI;electrospray ionization;EST;expressed sequence tag;HOP;heat shock protein (HSP)70/HSP90 organizing protein;HPLC;high-pressure liquid chromatography;IPG;immobilized pH gradient;LC;liquid chromatography;LL;constant conditions of dim light;MS;mass spectrometry;PDI;protein disulfide isomerase;S.E.M.;standard error of the mean;SDS;sodium dodecyl sulfate;TCA;trichloroacetic acid;TPR;tetratricopeptide repeat;WD-40 repeat;tandem repeats of about 40 residues;each containing a central Trp[W]-Asp[D] motif;2-DE;two-dimensional gel electrophoresis