Changes in kinesin distribution and phosphorylation occur during regulated secretion in pancreatic acinar cells.
Changes in kinesin distribution and phosphorylation occur during regulated secretion in pancreatic acinar cells.
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胰腺腺泡细胞的调节分泌过程中会发生驱动蛋白分布和磷酸化的变化。
DOI：
10.1016/s0171-9335(98)80056-3
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发表时间：
1998
影响因子：
6.6
通讯作者：
McNiven,MA
中科院分区：
文献类型：
--
作者：
Marlowe,KJ;Farshori,P;Torgerson,RR;Anderson,KL;Miller,LJ;McNiven,MA
在分泌细胞中，基于微管（Mt-）的运动酶被认为支持分泌囊泡运输到细胞表面并随后释放。目前，Mts和激酶在外分泌上皮细胞分泌囊泡运输中的作用尚未明确。此外，目前尚不清楚激动剂诱导的分泌事件是否会改变运动蛋白的功能和分布，从而改变囊泡运输。为此，我们利用分离的大鼠胰腺腺泡细胞和培养的大鼠胰腺腺泡细胞来检测Mts和激酶在调节分泌中的作用。细胞暴露于细胞骨架拮抗剂表明，观察到的顶端聚集的酶原颗粒（ZGs）的运动是由Mts支持的，而不是肌动蛋白。对极化腺泡中Mt +组织的形态学研究表明，Mt +末端从顶膜向细胞中心延伸。两种细胞模型的免疫荧光显微镜显示，激酶蛋白与根尖ZG明显相关，而胰腺亚细胞部分的定量免疫印迹分析证实，激酶蛋白在ZG膜上富集。此外，在培养的腺泡细胞中微量注射kinesin抗体可抑制ZG的运动。分离细胞的间接免疫荧光染色和分离ZGs的定量Western印迹显示，在分泌刺激下，激酶与颗粒膜的关联增加了3倍。32p标记的腺泡细胞放射自显影研究显示，在刺激分泌过程中，运动蛋白重链（KHC）磷酸化增加了6倍。这些研究提供了第一个直接证据，证明Mts和激酶支持ZG运动，生理激动剂诱导KHC显著磷酸化，同时增加激酶与ZG膜的关联。在激动剂刺激期间的这些变化表明，运动蛋白参与酶原分泌受到调节。
In secretory cells, microtubule- (Mt-) based motor enzymes are thought to support transport of secretory vesicles to the cell surface for subsequent release. At present, the role of Mts and kinesin in secretory vesicle transport in exocrine epithelial cells has not been defined. Furthermore, it is unclear whether an agonist-induced secretory event modifies kinesin function and distribution, thus altering vesicle transport. To this end, we utilized isolated rat pancreatic acini and cultured rat pancreatic acinar cells to examine the role of Mts and kinesin in regulated secretion. Exposure of cells to cytoskeletal antagonistic drugs demonstrated that the observed movements of apically clustered zymogen granules (ZGs) are supported by Mts, but not actin. Morphological studies of Mt organization in polarized acini show that Mt plus ends extend outward from the apical membrane toward the cell center. Immunofluorescence microscopy in both cell models revealed a clear association of kinesin with apical ZGs, while quantitative immunoblot analysis of pancreatic subcellular fractions confirmed kinesin enrichment on ZG membranes. In addition, microinjection of kinesin antibodies into cultured acinar cells inhibited ZG movements. Indirect immunofluorescence staining of isolated cells and quantitative Western blotting of isolated ZGs revealed that kinesin association with granule membranes increased up to 3-fold in response to a secretory stimulus. Autoradiographic studies of32P-labeled acini showed up to a 6-fold increase in kinesin heavy chain (KHC) phosphorylation during stimulated secretion. These studies provide the first direct evidence that Mts and kinesin support ZG movements and that physiological agonists induce a marked phosphorylation of KHC while increasing the association of kinesin with ZG membranes. These changes during agonist stimulation suggest that the participation of kinesin in zymogen secretion is regulated.
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嗜铬细胞中驱动蛋白的细胞内分布。
影响因子：
6.6
作者：
Schmitz,F;Wallis,KT;Rho,M;Drenckhahn,D;Murphy,DB
通讯作者：
Murphy,DB
DOI：
10.1083/jcb.113.5.1057
发表时间：
1991-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
Vitale ML;Rodríguez Del Castillo A;Tchakarov L;Trifaró JM
通讯作者：
Trifaró JM
培养细胞中细胞质动力蛋白免疫定位至溶酶体。
影响因子：
4
作者：
Sharron X. H. Lin;C. A. Collins
通讯作者：
C. A. Collins
神经递质释放过程中的细胞骨架动力学
影响因子：
15.9
作者：
Trifaró Jm;Vitale Ml
通讯作者：
Vitale Ml
环AMP通过增加单个细胞器运动的概率来调节海兔袋细胞神经元的快速轴突运输
影响因子：
16.2
作者：
E. Azhderian;D. Hefner;Chi;Leonard K. Kaczmarek;P. Forscher
通讯作者：
P. Forscher