The immune system) around the peripheral nerve fibers, which mainly consist of lymphocytes and monocytes (7, 55). In addition, in an animal model of the spontaneous autoimmune peripheral polyneuropathy (presymptomatic L31/CD4 / mice), the infiltration of immune cells was noted precisely in the areas of the DRG and spinal roots prior to disease onset (56). Thus, DRGs and dorsal roots theoretically
Produced in BL21 bacteria (Stratagene) using 0.5 mM IPTG induction. Bacterial pellets were lysed in NETN buffer (50 mM Tris-HCl pH8.0, 150 mM NaCl, 0.5 NP40, 5 mM EDTA, PIs), cleared by ultracentrifugation and bound to glutathione sepharose 4B (GE Healthcare) for 1 hr at 4 . Beads were washed with NETN, and amount of GST protein was estimated by SDS-PAGE followed by Coomassie staining. GST fusion
Produced in BL21 bacteria (Stratagene) using 0.5 mM IPTG induction. Bacterial pellets were lysed in NETN buffer (50 mM Tris-HCl pH8.0, 150 mM NaCl, 0.5 NP40, 5 mM EDTA, PIs), cleared by ultracentrifugation and bound to glutathione sepharose 4B (GE Healthcare) for 1 hr at 4 . Beads were washed with NETN, and amount of GST protein was estimated by SDS-PAGE followed by Coomassie staining. GST fusion
Produced in BL21 bacteria (Stratagene) using 0.5 mM IPTG induction. Bacterial pellets were lysed in NETN buffer (50 mM Tris-HCl pH8.0, 150 mM NaCl, 0.5 NP40, 5 mM EDTA, PIs), cleared by ultracentrifugation and bound to glutathione sepharose 4B (GE Healthcare) for 1 hr at 4 . Beads were washed with NETN, and amount of GST protein was estimated by SDS-PAGE followed by Coomassie staining. GST fusion
Y meaningful that proteins such as ESP15 (a type I TM protein that localized to the lysosome) is in the surface proteome, whereas p67 (also a type I TM glycoprotein) is not. Membrane Domains and Domain Maintenance--The few surface proteins analyzed to date suggest the existence of at least three biochemically distinct domains across contiguous membranes, and emphasize the idea that individual prot
An NMR spectroscopist, frequently raise the following question: the structure of a homologous protein has been resolved by X-raycrystallography, does this ease the resonance assignment of my protein? The answer is unfortunately negative. Numerous effects control the NMR chemical shifts and thus, even for a protein with a known structure, it is nearly impossible to predict them. In other terms, the
An NMR spectroscopist, frequently raise the following question: the structure of a homologous protein has been resolved by X-raycrystallography, does this ease the resonance assignment of my protein? The answer is unfortunately negative. Numerous effects control the NMR chemical shifts and thus, even for a protein with a known structure, it is nearly impossible to predict them. In other terms, the
Harvested and resuspended in a lysis buffer containing 0.4 M sucrose, 50 mM 3-(Nmorpholino) propanesulfonic acid, pH 7.0, 10 mM NaCl, 5 mM EDTA, and 0.5 mM PMSF. Cells were broken using a bead beater, and centrifuged with 5,000 g for 30 min at 4 to remove glass beads and insoluble cell debris. The membrane and the soluble fractions were separated as previously described with slight modifications