As used to dry them prior to resuspension in 5 (v/v) formic acid. Peptide concentrations were determined using the CBQCA assay after 25-fold dilution of peptide samples in 0.1 M borate buffer, pH 9.3. Denaturing Size-exclusion Chromatography--For the denaturing SEC, cells were lysed in PBS containing inhibitors as above and supplemented with 4 SDS, 25 mM TCEP, and 50 mM N-ethylmaleimide. The col
Ying SNARE complex assembly and disassembly and their control by chaperones including membrane tethering factors, Sec1-Munc18 proteins (SM proteins) [G], and Nethylmaleimide-sensitive fusion factor (NSF) [G]. Membrane tethering factors8-10 not only mediate the initial contact between a vesicle and its target membrane but also bind SNAREs to regulate their assembly. Tethering factors work together
Tant of E. coli growing under ammonium-limited conditions accumulated glutamine (12 mM) relative to the wild type (2 mM) (135). Induction of AmtB from a plasmid by IPTG (in order to increase the intracellular ammonium concentration so that GDH would be active) did not have the anticipated positive effect on growth of the gltB mutant, however (281). (v) The glnHPQ operon. E. coli can use glutamine
Tant of E. coli growing under ammonium-limited conditions accumulated glutamine (12 mM) relative to the wild type (2 mM) (135). Induction of AmtB from a plasmid by IPTG (in order to increase the intracellular ammonium concentration so that GDH would be active) did not have the anticipated positive effect on growth of the gltB mutant, however (281). (v) The glnHPQ operon. E. coli can use glutamine