TY - JOUR
TI - Analysis of Water-Soluble Proteins by Two-Dimensional Electrophoresis in the Encystment Process of Colpoda cucullus Nag-1 and Cytoskeletal Dynamics
AU - Sogame, Yoichiro
AU - Kojima, Katsuhiko
AU - Takeshita, Toshikazu
AU - Kikuchi, Shiho
AU - Shimada, Yuto
AU - Nakamura, Rikiya
AU - Arikawa, Mikihiko
AU - Miyata, Seiji
AU - Kinoshita, Eiji
AU - Suizu, Futoshi
AU - Matsuoka, Tatsuomi
TI - Analysis of Water-Soluble Proteins by Two-Dimensional Electrophoresis in the Encystment Process of Colpoda cucullus Nag-1 and Cytoskeletal Dynamics
AB - 
	Assays of protein contained in water-soluble fraction of encysting cells Colpoda cucullus Nag-1 by two-dimensional electrophoresis (2-D PAGE) and mass spectrometry (MS) revealed that the amount of β-tubulin abruptly increased in 2.5–10 h after encystment induction. Judging from the results that total α-tubulin content did not decrease much until 12 h after encystment induction, the result indicates that disassembly of microtubules may occur soon after encystment is induced. Therefore, we tried to visualize dynamics of microtubules. Immunofluorescence microscopy using anti-α-tubulin antibody indicated that disassembly of axonemal microtubules of cilia became within 1.5 h after encystment induction, and resorbed in 3 days. Although the cytoplasmic microtubules failed to be visualized clearly, encystmentdependent globulation of cells was promoted by taxol, an inhibitor of disassembly of microtubules. It is possible that a temporary formation of cytoplasmic microtubules may be involved in cell globulation.

	The phosphorylation level of actin (43 kDa) became slightly elevated just after encystment induction. Lepidosomes, the sticky small globes surrounding encysting cells, were vividly stained with Acti-stain 555 phalloidin, suggesting that 43-kDa actin or its homologues may be contained in lepidosomes.
VL - 2020
IS - Volume 59, Issue 3-4
PY - 2021
SN - 0065-1583
C1 - 1689-0027
SP - 107
EP - 120
DO - 10.4467/16890027AP.20.009.13264
UR - https://ejournals.eu/en/journal/acta-protozoologica/article/analysis-of-water-soluble-proteins-by-two-dimensional-electrophoresis-in-the-encystment-process-of-colpoda-cucullus-nag-1-and-cytoskeletal-dynamics
KW - resting cyst formation
KW - microtubules
KW - actin
KW - lepidosomes
KW - resorption of cilia.