Detail of "9035-75-0"

Reference

Identification of cDNA clones encoding secretory isoenzyme forms: Sequence determination of canine pancreatic prechymotrypsinogen 2 mRNA

Identification of cDNA clones encoding secretory isoenzyme forms: Sequence determination of canine pancreatic prechymotrypsinogen 2 mRNA. Pinsky, Seth D.; LaForge, K. Steven; Luc, Van; Scheele, George (Lab. Cell Mol. Biol., Rockefeller Univ., New York, NY 10021, USA). Proc. Natl. Acad. Sci. U. S. A., 80(24), 7486-90 (English) 1983. CODEN: PNASA6. ISSN: 0027-8424. DOCUMENT TYPE: Journal CA Section: 3 (Biochemical Genetics) Section cross-reference(s): 13 A cDNA library has been constructed from canine poly(A)+ mRNA. Clones contg. cDNA inserts coding for prechymotrypsinogen 2 (isoelec. point = 7.1; mol. wt. = 27,500), one of 3 canine pancreatic isoenzyme forms, were selected by colony hybridization using a cDNA probe synthesized from immunoselected prechymotrypsinogen 2 mRNA. To verify that cDNA clones code for prechymotrypsinogen 2 forms that translocate across rough endoplasmic reticulum membranes and fold into stable and identifiable secretory proteins, in vitro translation of hybrid-selected mRNA was conducted in the presence of microsomal membranes and optimal concns. of glutathione and nascent translation products were analyzed in their nonreduced state by 2-dimensional isoelec. focusing/SDS-gel electrophoresis and fluorog. A near full-length chymotrypsinogen 2 (EC 3.4.21.1) [9035-75-0] cDNA and its primed extension were used to det. the nucleotide sequence for the entire coding region of prechymotrypsinogen 2 mRNA and 87 residues, including a poly(A) addn. signal, in the 3' nontranslated region. The deduced amino acid sequence shows a 263-residue presecretory protein contg. an 18-residue N-terminal transport peptide, which has been previously shown to mediate the translocation of chymotrypsinogen 2 across the rough endoplasmic reticulum membrane. 89190-70-5 and 89190-72-7 are just another two chemicals used in this study. Following the transport peptide is a 245-residue proenzyme, which shows 82% and 80% sequence identity with bovine chymotrypsinogens A and B, resp. Conserved among the 3 zymogens are 10 Cys residues that form 5 SS bonds in bovine chymotrypsinogens A and B and the residues that are required for zymogen activation, substrate binding, and catalytic activity. .

Electrophoretic extraction-concentration of proteins from polyacrylamide gels under alkaline, neutral and acidic conditions

Electrophoretic extraction-concentration of proteins from polyacrylamide gels under alkaline, neutral and acidic conditions. Hashizume, Shuichi; Rashid, Mohammad A.; Shoji, Masahiro; Kuroda, Kazuhiko (Morinaga Inst. Biol. Sci., Yokohama 230, Japan). Electrophoresis (Weinheim, Fed. Repub. Ger.), 5(1), 30-4 (English) 1984. CODEN: ELCTDN. ISSN: 0173-0835. DOCUMENT TYPE: Journal CA Section: 9 (Biochemical Methods) A simple technique was developed for the electrophoretic extn. of milligram to microgram quantities of protein, under const. monitoring by dye line, in a concd. form from polyacrylamide gels run under alk., neutral and acidic conditions. Typically, the extn. time from 5 gel slices per tube contg. 9068-76-2 and 9035-75-0 which are cas registry numbers are also used here. ~1.2 mg bovine serum albumin or a-chymotrypsinogen A was 3 h, and the max. concn. of the recovered protein was 5 mg/mL with a total recovery of >90% in all cases. Higher protein concn. (11 mg/mL) could be obtained by increasing the extn. time to 3.5 h or by using resins like glass beads. The alk. extn. system was applied to the purifn. of glutamyl-tRNA synthetase up to homogeneity, and also to ferritin. This technique can also be used under reducing conditions and the scale of operation can be adjusted by the no. of tubes or no. of slices. .