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首页> 外文期刊>Molecular biology of the cell >A Cytosolic Splice Variant of Cab45 Interacts with Munc18b and Impacts on Amylase Secretion by Pancreatic Acini
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A Cytosolic Splice Variant of Cab45 Interacts with Munc18b and Impacts on Amylase Secretion by Pancreatic Acini

机译：Cab45的胞质剪接变体与Munc18b相互作用，并影响胰腺腺泡蛋白分泌淀粉酶。

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摘要

We identified in a yeast two-hybrid screen the EF-hand Ca2+-binding protein Cab45 as an interaction partner of Munc18b. Although the full-length Cab45 resides in Golgi lumen, we characterize a cytosolic splice variant, Cab45b, expressed in pancreatic acini. Cab45b is shown to bind 45Ca2+, and, of its three EF-hand motifs, EF-hand 2 is demonstrated to be crucial for the ion binding. Cab45b is shown to interact with Munc18b in an in vitro assay, and this interaction is enhanced in the presence of Ca2+. In this assay, Cab45b also binds the Munc18a isoform in a Ca2+-dependent manner. The endogenous Cab45b in rat acini coimmunoprecipitates with Munc18b, syntaxin 2, and syntaxin 3, soluble N -ethylmaleimide-sensitive factor attachment protein receptors with key roles in the Ca2+-triggered zymogen secretion. Furthermore, we show that Munc18b bound to syntaxin 3 recruits Cab45b onto the plasma membrane. Importantly, antibodies against Cab45b are shown to inhibit in a specific and dose-dependent manner the Ca2+-induced amylase release from streptolysin-O–permeabilized acini. The present study identifies Cab45b as a novel protein factor involved in the exocytosis of zymogens by pancreatic acini.
机译：我们在酵母双杂交筛选中确定EF手Ca 2 + 结合蛋白Cab45为Munc18b的相互作用伴侣。虽然全长Cab45驻留在高尔基体腔中，但我们表征了在胰腺腺泡中表达的胞质剪接变体Cab45b。 Cab45b已显示与 45 Ca 2 + 结合，并且在其三个EF-手基序中，EF-hand 2对离子结合至关重要。 Cab45b在体外试验中显示与Munc18b相互作用，并且在Ca 2 + 存在下这种相互作用得以增强。在该测定中，Cab45b还以Ca 2 + 依赖的方式结合Munc18a同工型。大鼠acini中的内源性Cab45b与Munc18b，syntaxin 2和syntaxin 3，可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体共沉淀，在Ca 2 + 触发的酶原分泌中起关键作用。此外，我们显示绑定到syntaxin 3的Munc18b将Cab45b募集到质膜上。重要的是，已显示出抗Cab45b的抗体以特异性和剂量依赖性的方式抑制Ca 2+诱导的淀粉酶从链球菌溶血素-O-通透的腺泡中释放。本研究确定Cab45b是一种新型蛋白因子，参与胰腺腺泡对酶原的胞吐作用。

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