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Switching of cargo sorting from the constitutive to regulated secretory pathway by the addition of cystatin D sequence in salivary acinar cells

Am J Physiol Gastrointest Liver Physiol. 2020; 
Junko Fujita-Yoshigaki, Megumi Yokoyama, Osamu Katsumata-Kato
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Insect Expression … described previously(40, 41). The construct with FLAG tag at N-terminal and 10 × His at carboxyl terminal was synthesized by GenScript and then cloned into pFastBac1 vector and was expressed in Spodoptera frugiperda Sf9 insect cells using the baculovirus method … Get A Quote

摘要

The mechanism for segregation of cargo proteins into the regulated and constitutive secretory pathways in exocrine cells remains to be elucidated. We examined the transport of HaloTag proteins fused with full-length cystatin D (fCst5-Halo) or only its signal peptide (ssCst5-Halo) in parotid acinar cells. Although both fusion proteins were observed to be colocalized with amylase in the secretory granules, the coefficients for overlapping and correlation of fCst5-Halo with amylase were higher than those of ssCst5-Halo. The secretion of both the proteins was enhanced by the addition of the β-adrenergic receptor agonist isoproterenol as well as endogenous amylase. In contrast, unstimulated secretion of ssCst5-Halo... More

关键词

computer simulation, exocrine glands, protein transport, secretory granule, sorting signals