Using periodate-oxidized nucleotide as affinity label for the nucleotide site of proteins
- 1 October 1993
- journal article
- Published by Springer Nature in Protein Journal
- Vol. 12 (5) , 627-632
- https://doi.org/10.1007/bf01025127
Abstract
The active site of pigeon liver malic enzyme was labeled with a fluorescent affinity label, the periodate-oxidized aminopyridine adenine dinucleotide phosphate. The modified enzyme was subjected to proteolytic digestion with trypsin. The resulted peptides were then separated with reversed-phase high-performance liquid chromatography on Waters μBondapak C18 column. Two pure fluorescent peptides were obtained after three runs of the chromatography. The peptides were then subjected to automatic Edman degradation on a Beckman peptide sequencer and subsequently separated and identified with phenylthiohydantoin C18 column. No sequence was obtained. The possible reasons for the failure in sequencing the periodate-oxidized nucleotides labeled active site peptide and some possible pitfalls in using these reagents were discussed.Keywords
This publication has 23 references indexed in Scilit:
- Modification of the ATP inhibitory site of the Ascaris suum phosphofructokinase results in the stabilization of an inactive T stateBiochemistry, 1991
- Duck liver malic enzyme: sequence of a tryptic peptide containing the cysteine residue labeled by the substrate analog bromopyruvateBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1991
- Oxidation and cleavage of 3-aminopyridine adenine dinucleotide phosphate with periodateJournal of Biochemical and Biophysical Methods, 1990
- Structural identity of the subunits of pigeon liver malic enzymeInternational Journal of Peptide and Protein Research, 1987
- Isolation and sequence determination of an active site peptide of rabbit muscle pyruvate kinaseArchives of Biochemistry and Biophysics, 1987
- Affinity labeling of nicotinamide adenine dinucleotide-dependent isocitrate dehydrogenase by the 2',3'-dialdehyde derivative of adenosine 5'-diphosphate. Evidence for the formation of an unusual reaction productBiochemistry, 1983
- Interactions between the mitochondrial adenosinetriphosphatase and periodate-oxidized adenosine 5'-triphosphate, an affinity label for adenosine 5'-triphosphate binding sitesBiochemistry, 1982
- Oxidized NADP as a potential active-site-directed reagent of pigeon liver malic enzymeBiochemical and Biophysical Research Communications, 1979
- Affinity labeling of bovine colostrum galactosyltransferase with a uridine 5'-diphosphate derivativeBiochemistry, 1976
- Identification of the chemical groups involved in the binding of periodate-oxidized NADP+ to 6-phosphogluconate dehydrogenaseBiochimica et Biophysica Acta (BBA) - Enzymology, 1976