High-performance liquid affinity chromatography and in situ fluorescent labeling on thin-layer chromatography of glycosphingolipids.
AuthorsTomono Y, Abe K, Watanabe K
JournalAnal Biochem
PubMed ID2327579
'A method combining high-performance liquid affinity chromatography and in situ fluorescent labeling on thin-layer chromatography is introduced for determination of glycosphingolipids. Glycolipids in crude extract from rat liver were separated quantitatively from neutral lipids and phospholipids with a phenylboronic acid-derivatized silica gel column. Glycolipids were eluted quantitatively with approximately 98% ... More
Visual detection of peptidase activity using fluorogenic substrates in a microtiter plate assay.
AuthorsIrvine GB, Ennis M, Williams CH
JournalAnal Biochem
PubMed ID2339787
'A simple, inexpensive, and sensitive assay for peptidase activity has been devised. The assay was performed in a microtiter plate and was based on fluorogenic peptide substrates, many of which are commercially available. 7-Amino-4-methyl coumarin the fluorescent product liberated during an incubation period of between 1 and 16 h, was ... More
New fluorogenic substrates for subtilisin.
AuthorsKanaoka Y, Takahashi T, Nakayama H, Tanizawa K
JournalChem Pharm Bull (Tokyo)
PubMed ID3899387
A new fluorescent tag for labeling of saccharides.
AuthorsPrakash C, Vijay IK
JournalAnal Biochem
PubMed ID6846798
Potential aldehyde groups of N-acetylglucosamine and its di- and tri-saccharides were coupled with 7-amino-4-methylcoumarin (AMC) by reductive amination in the presence of sodium cyanoborohydride. The products were purified by silica gel chromatography. Acid hydrolysis and digestion with glycosidases gave the expected products. One picomole of the sugar-AMC complexes could be ... More
A new fluorogenic substrate for plasmin.
AuthorsPierzchala PA, Dorn CP, Zimmerman M
JournalBiochem J
PubMed ID161507
A new fluorogenic peptide substrate for plasmin, 7-(N-succinoylalanylphenylalanyl-lysylamido)-4-methylcoumarin trifluoroacetate salt, was prepared that can be used in a simple and direct assay. The results obtained by the assay method are linear over a wide range of enzyme concentrations and sensitive enough to detect as little as 10(-5) CTA units of ... More
New intramolecularly quenched fluorogenic peptide substrates for the study of the kinetic specificity of papain.
AuthorsGarcía-Echeverría C, Rich DH
JournalFEBS Lett
PubMed ID1551413
A series of new substrates for determining the catalytic activity of cysteine proteinases is described. The rate of hydrolysis by papain was monitored by a fluorescence continuous assay based on internal resonance energy transfer using 5-[(2-aminoethyl)amino]naphtalene-1-sulfonic acid (EDANS) and 4-(4-dimethylaminophenylazo)benzoic acid (DABCYL) as fluorescent donor and quenching acceptor, respectively, in ... More
Effect of N-methylation of phosphatidylethanolamine on the fluidity of phospholipid bilayers.
AuthorsMio M, Okamoto M, Akagi M, Tasaka K
JournalBiochem Biophys Res Commun
PubMed ID6732794
The effect of N-methylphosphatidylethanolamine on phase transition and the fluidity of the liposomes made of dipalmitoylphosphatidylcholine or phosphatidylethanolamine was studied by the steady-state fluorescence polarization method and differential scanning calorimetry. N-methylation of phosphatidylethanolamine caused a decrease of fluidity of liposomes made of dipalmitoylphosphatidylcholine, but had little effect on dipalmitoylphosphatidylethanolamine. The ... More
Studies on immunoassays of peptide factors. II. Fluorescence enzyme immunoassay for human little-gastrin.
AuthorsMoroder L, Bovermann G, Mourier G, Göhring W, Gemeiner M, Wünsch E
JournalBiol Chem Hoppe Seyler
PubMed ID3304342
The fluorogenic chymotrypsin substrate N alpha-(4-carboxybutyryl)-L-phenylalanine (4-methyl-7-coumaryl)amide was converted to a thiol-containing compound via its condensation at the carboxyl function with cystamine followed by reduction of the resulting disulfide compound to a cysteamine derivative. By subsequent reaction of the thiol group with N alpha-maleoyl-beta-alanyl-human-little-gastrin-I-[2-17] a fluorogenic substrate-labeled gastrin, fully immunoreactive ... More
Ultraviolet B radiation generates platelet-activating factor-like phospholipids underlying cutaneous damage.
AuthorsMarathe GK, Johnson C, Billings SD, Southall MD, Pei Y, Spandau D, Murphy RC, Zimmerman GA, McIntyre TM, Travers JB
JournalJ Biol Chem
PubMed ID16115894
Ultraviolet B light (UVB) causes cutaneous inflammation and cell death, but the agents responsible are not defined. These studies examined the role of the platelet-activating factor (PAF) signaling system in UVB-mediated effects. Expression of the PAF receptor in the PAF receptor-negative epidermoid cell line KB augmented apoptosis in response to ... More
Interactions of phage P22 tails with their cellular receptor, Salmonella O-antigen polysaccharide.
AuthorsBaxa U, Steinbacher S, Miller S, Weintraub A, Huber R, Seckler R
JournalBiophys J
PubMed ID8889178
Bacteriophage P22 binds to its cell surface receptor, the repetitive O-antigen structure in Salmonella lipopolysaccharide, by its six homotrimeric tailspikes. Receptor binding by soluble tailspikes and the receptor-inactivating endorhamnosidase activity of the tailspike protein were studied using octa- and dodecasaccharides comprising two and three O-antigen repeats of Salmonella enteritidis and ... More
Cytochemistry of membrane proteases.
AuthorsGossrau R
JournalHistochem J
PubMed ID3902743
Membrane proteases that are detectable by cytochemical means are the classified exopeptidases, aminopeptidases A and M (or N), gamma-glutamyl transpeptidase (which also acts as transferase), dipeptidyl peptidase IV and the endopeptidase, enteropeptidase (also known as enterokinase). Not yet classified are the possible exopeptidase, tripeptidyl peptidase and endopeptidases I (Ala-endopeptidase) and ... More