Quantitation of platelet-activating factor by high-performance liquid chromatography with fluorescent detection.
AuthorsMita H, Yasueda H, Hayakawa T, Shida T
JournalAnal Biochem
PubMed ID2817335
'Platelet-activating factors, 1-O-hexadecyl- and 1-O-octadecyl-2-acetyl-sn-glycero-3-phosphocholine (C16-AGEPC and C18AGEPC), were measured by reverse-phase high-performance liquid chromatography with fluorescent detection. C16AGEPC, C18AGEPC, and 1-O-hexadecyl-2-propionyl-sn-glycero-3-phosphocholine, which was suitable for use as an internal standard, were hydrolyzed with phospholipase C, and then the resulting hydrolyzed products were derivatized with 7-methoxycoumarin-3-carbonyl chloride or 7-methoxy-coumarin-4-acetic acid ... More
An alternative quenched fluorescence substrate for Pz-peptidase.
AuthorsTisljar U, Knight CG, Barrett AJ
JournalAnal Biochem
PubMed ID2192576
7-Methoxycoumarin-3-carboxylyl-Pro-Leu-Gly-Pro-D-Lys(2,4-dinitr oph enyl) is introduced as a new quenched fluorescence substrate for assaying Pz-peptidase (also known as soluble metallo-endopeptidase and endo-oligopeptidase). The value of Km for partially purified Pz-peptidase from rat muscle was 8.6 microM. High protein concentrations did not interfere with the assay, so that for the first time ... More
A modular strategy for tailoring fluorescent biosensors from ribonucleopeptide complexes.
AuthorsHagihara M, Fukuda M, Hasegawa T, Morii T
JournalJ Am Chem Soc
PubMed ID17002390
Fluorescent biosensors that facilitate reagentless sensitive detection of small molecules are crucial tools in the areas of therapeutics and diagnostics. However, construction of fluorescent biosensors with desired characteristics, that is, detection wavelengths and concentration ranges for ligand detection, from macromolecular receptors is not a straightforward task. An ATP-binding ribonucleopeptide (RNP) ... More