[A comparative level of expression of some proteins in XL2 cell synchronized on different phases of cell cycle].
Cells of cultured line XL2 (Xenopus laevis) were synchronized by a combine effect of serum deprivation, aphidicolin, nocodazole and ALLN treatments. Four fractions were prepared, with maximum percentage of cells being in G1, S and G2 phases of cell cycle, and in mitosis, respectively. Comparative levels of six different proteins (beta-tubulin, DNA topoisomerase IIa, Xenopus Aurora A kinase pEg2, kinesin-like motor protein X1Eg5, and two members of condensis family proteins pEg7 (XCAP D2) and XCAP E were detected by quantitative Western blot analysis of these fractions. We used a new method of mathematic processing of data that commonly provides a possibility to calculate a comparative quantity of proteins in hypothetically "clean" fraction composed of cells being in the same phase of the cell cycle. This method makes it possible to use even partly synchronized cell cultures for analysis of changes in protein quantity, provided a precede determination of cell population composition is made.
Uzbekov
R E
RE
Division of Electron Microscopy, A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University. rustuzbekov@aol.com
D000818:Animals; D064127:Aurora Kinases; D015153:Blotting, Western; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000096:biosynthesis*; D002460:Cell Line; D004250:DNA Topoisomerases, Type II / Q000096:biosynthesis*; D004527:Egg Proteins / Q000096:biosynthesis*; D011494:Protein Kinases / Q000096:biosynthesis*; D017346:Protein Serine-Threonine Kinases; D014404:Tubulin / Q000096:biosynthesis*; D029867:Xenopus Proteins / Q000096:biosynthesis* / Q000378:metabolism*; D014982:Xenopus laevis
D018797:Cell Cycle Proteins; C116601:Eg7 protein, Xenopus; D004527:Egg Proteins; D014404:Tubulin; D029867:Xenopus Proteins; D011494:Protein Kinases; C112134:AURKA protein, Xenopus; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases; D004250:DNA Topoisomerases, Type II