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441.
442.
Pancreatic acinar cells produce and secrete digestive enzymes. These cells are organized as a cluster which forms and shares a joint lumen. This work demonstrates how the secretory capacity of these cells can be assessed by culture of isolated acini. The setup is advantageous since isolated acini, which retain many characteristics of the intact exocrine pancreas can be manipulated and monitored more readily than in the whole animal. Proper isolation of pancreatic acini is a key requirement so that the ex vivo culture will represent the in vivo nature of the acini. The protocol demonstrates how to isolate intact acini from the mouse pancreas. Subsequently, two complementary methods for evaluating pancreatic secretion are presented. The amylase secretion assay serves as a global measure, while direct imaging of pancreatic secretion allows the characterization of secretion at a sub-cellular resolution. Collectively, the techniques presented here enable a broad spectrum of experiments to study exocrine secretion. 相似文献
443.
Daniel Ronen Masha M. Rosenberg Deborah E. Shalev Michael Rosenberg Shahar Rotem Assaf Friedler Shoshana Ravid 《The Journal of biological chemistry》2010,285(10):7079-7086
The motor protein, non-muscle myosin II (NMII), must undergo dynamic oligomerization into filaments to participate in cellular processes such as cell migration and cytokinesis. A small non-helical region at the tail of the long coiled-coil region (tailpiece) is a common feature of all dynamically assembling myosin II proteins. In this study, we investigated the role of the tailpiece in NMII-C self-assembly. We show that the tailpiece is natively unfolded, as seen by circular dichroism and NMR experiments, and is divided into two regions of opposite charge. The positively charged region (Tailpiece1946–1967) starts at residue 1946 and is extended by seven amino acids at its N terminus from the traditional coiled-coil ending proline (Tailpiece1953–1967). Pull-down and sedimentation assays showed that the positive Tailpiece1946–1967 binds to assembly incompetent NMII-C fragments inducing filament assembly. The negative region, residues 1968–2000, is responsible for NMII paracrystal morphology as determined by chimeras in which the negative region was swapped between the NMII isoforms. Mixing the positive and negative peptides had no effect on the ability of the positive peptide to bind and induce filament assembly. This study provides molecular insight into the role of the structurally disordered tailpiece of NMII-C in shifting the oligomeric equilibrium of NMII-C toward filament assembly and determining its morphology. 相似文献
444.
Lea Eisenbach Ofer Mandelboim Erez Bar-Haim Lior Carmon Hernan Copcow Khaled El-Shami Adrian Paz Dan Popovic Ezra Vadai Esther Tzehoval Michael Feldman Mati Fridkin 《Letters in Peptide Science》1998,5(5-6):323-328
Cytotoxic T-lymphocytes (CTLs) kill abnormal cells. CTLs recognize major histocompatibility complex class I molecules in complex with peptides derived from relevant antigens. The identification of tumor associated antigen peptides enabled the design of anti-tumor and anti-metastatic vaccines in a murine lung carcinoma. 相似文献