In vivo calcineurin crystals formed using the baculovirus expression system
| Autor: | G Y, Fan, F, Maldonado, Y, Zhang, R, Kincaid, M H, Ellisman, L N, Gastinel |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Cell Nucleus
Recombination Genetic Cytoplasm Insecta Neurospora crassa Calcineurin Genetic Vectors Polymerase Chain Reaction Gene Expression Regulation Enzymologic Recombinant Proteins Microscopy Electron Phosphoprotein Phosphatases Animals Humans Calmodulin-Binding Proteins Electrophoresis Polyacrylamide Gel Crystallization Baculoviridae Cells Cultured |
| Zdroj: | Microscopy research and technique. 34(1) |
| ISSN: | 1059-910X |
| Popis: | Calcineurin is a heterodimeric phosphatase involved in the signal transduction of antigen-activated T cells. Coexpression of its two subunits, the regulatory subunit from human and the catalytic subunit from Neurospora crassa in cultured insect cells using the baculovirus expression system results in the formation of very large crystals in the cytoplasm. The crystals are formed initially in vesicles, but their subsequent growth appears to be uninhibited and continues without the need of an enclosing membrane until the host cell lyses. Although these in vivo crystals are low in population, ranging only 0-3 per cell, they are extremely large, over 10 mu m in some cases. Biochemical assays confirm their calcineurin origin, with the regulatory subunit incorporated being myristoylated, although both the myristoylated and unmyristoylated forms are expressed. The lattice structure of the in vivo crystals, with a spacing of 5.5 nm, is preserved with the regular electron microscopic (EM) specimen preparation procedure. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text