Nuclear calcium flux in Trypanosoma brucei can be quantified with targeted aequorin
作者： Zhao-Hui Xiong , Larry Ruben
DOI: 10.1016/S0166-6851(96)02750-8
关键词: Calcium 、 Aequorin 、 Molecular biology 、 Cytoplasm 、 Nucleus 、 Calcium channel 、 Biology 、 Calcium flux 、 Photoprotein 、 Ionomycin 、 Parasitology
摘要: The following study was undertaken to determine if calcium ions move from the plasma membrane nucleus of Trypanosoma brucei. Nuclear and cytosolic flux measured with sensitive photoprotein, aequorin which targeted various locations in stably transformed procyclic cells. Immunoblots revealed that recombinant proteins, CYT-AEQ NUC-AEQ were translated transformants, contained a soluble fraction. Immunolocalization demonstrated within trypanosome nucleus. To evaluate movement live trypanosomes, reconstituted vivo coelenterazine luminescence recorded. resting levels [Ca2+]cyt [Ca2+]nuc similar (314 ± 43 287 28 nM, respectively). When influx across initiated 2 μM ionomycin, each became elevated parallel new steady state approximately 2-fold above level. Compound 4880 by different mechanism manner inhibited channel antagonist, La3+. (8 μg/ml) transiently 1.73 0.3 over course 20 s, also generated transient rise peaked at 1.32 + 0.29 same time course. Overall, these data demonstrate moves into out closely parallels changes [Ca2+]cyt. A small ion gradient between cytoplasm observed.
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