1.Development of a biotinylated analog of substance P for use as a receptor probe.
Anton PA1, Reeve JR Jr, Vidrich A, Mayer E, Shanahan F. Lab Invest. 1991 May;64(5):703-8.
The development of a biotinylated substance P (SP) analog for use as a receptor probe is reported. The lysine in position 3 of SP was substituted by arginine and an amino terminal extension (NTE-SP) was added consisting of Lys-Tyr-Gly-Gly-Gly-Gly-Gly-Gly. Biotinylation of the N-terminal lysine was performed. The biotinylated peptide was purified by high performance liquid chromatography and characterized by mass spectral analysis. Binding studies using human IM-9 lymphoblasts with the biotinylated SP analog (biotin-NTE[Arg3]SP) and native SP yielded dissociation curves which were identical. In addition, the biotinylated SP analog retained functional activity similar to that of native SP in altering intracellular calcium concentration of Fura-2 loaded isolated rabbit colonic myocytes. Applicability of the SP receptor probe was demonstrated by using the streptavidin-peroxidase detection system to identify SP receptors on human IM-9 lymphoblasts.
2.Novel N-terminal sequence of a glycine-rich protein in the aleurone layer of soybean seeds.
Matsui M1, Toyosawa I, Fukuda M. Biosci Biotechnol Biochem. 1994 Oct;58(10):1920-2.
Glycine-rich protein (GRP), a cell wall protein, was extracted with hot water from the aleurone layer of soybean seeds. GRP was purified by adsorption on DEAE-Sephadex, Sephadex G-100 gel chromatography, and anion exchange HPLC. The estimated molecular size of GRP was approximately 30 kDa and GRP contained 59% glycine and 15% serine. The N-terminal amino acid sequence was a novel Gly-Gly-Gly-Gly-Gly-Gly-Gly-Gly-Gly-Gly-Gly-Gly-Gly-Gly-Gly-.
