title: Inactivation of brain myo-inositol monophosphate phosphatase by pyridoxal-5'-phosphate.
abstract: Myo-inositol monophosphate phosphatase (IMPP) is a key enzyme in the phosphoinositide cell-signaling system. This study found that incubating the IMPP from a porcine brain with pyridoxal-5'-phosphate (PLP) resulted in a time-dependent enzymatic inactivation. Spectral evidence showed that the inactivation proceeds via the formation of a Schiff's base with the amino groups of the enzyme. After the sodium borohydride reduction of the inactivated enzyme, it was observed that 1.8 mol phosphopyridoxyl residues per mole of the enzyme dimer were incorporated. The substrate, myo-inositol-1-phosphate, protected the enzyme against inactivation by PLP. After tryptic digestion of the enzyme modified with PLP, a radioactive peptide absorbing at 210 nm was isolated by reverse-phase HPLC. Amino acid sequencing of the peptide identified a portion of the PLP-binding site as being the region containing the sequence L-Q-V-S-Q-Q-E-D-I-T-X, where X indicates that phenylthiohydantoin amino acid could not be assigned. However, the result of amino acid composition of the peptide indicated that the missing residue could be designated as a phosphopyridoxyl lysine. This suggests that the catalytic function of IMPP is modulated by the binding of PLP to a specific lysyl residue at or near its substrate-binding site of the protein.
authors.affiliation: Department of Genetic Engineering and Research Institute for Bioscience and Biotechnology, Hallym University, Chunchon 200-702, Korea.
authors.forename: Dae Won
authors.initials: DW
authors.lastname: Kim
authors.forename: Joung Woo
authors.initials: JW
authors.lastname: Hong
authors.forename: Won Sik
authors.initials: WS
authors.lastname: Eum
authors.forename: Hee Soon
authors.initials: HS
authors.lastname: Choi
authors.forename: Soo Hyun
authors.initials: SH
authors.lastname: Choi
authors.forename: So Young
authors.initials: SY
authors.lastname: Kim
authors.forename: Byung Ryong
authors.initials: BR
authors.lastname: Lee
authors.forename: Jae Jin
authors.initials: JJ
authors.lastname: An
authors.forename: Sun Hwa
authors.initials: SH
authors.lastname: Lee
authors.forename: Seung Ree
authors.initials: SR
authors.lastname: Lee
authors.forename: Oh-Shin
authors.initials: OS
authors.lastname: Kwon
authors.forename: Hyeok Yil
authors.initials: HY
authors.lastname: Kwon
authors.forename: Sung-Woo
authors.initials: SW
authors.lastname: Cho
authors.forename: Kil Soo
authors.initials: KS
authors.lastname: Lee
authors.forename: Jinseu
authors.initials: J
authors.lastname: Park
authors.forename: Soo Young
authors.initials: SY
authors.lastname: Choi
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D001665:Binding Sites; D001894:Borohydrides / Q000494:pharmacology; D001921:Brain / Q000201:enzymology*; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D004789:Enzyme Activation / Q000187:drug effects; D007295:Inositol Phosphates / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000378:metabolism; D010669:Phenylthiohydantoin / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011485:Protein Binding; D011732:Pyridoxal Phosphate / Q000378:metabolism / Q000494:pharmacology*; D013552:Swine; D013997:Time Factors
chemical_list: D000596:Amino Acids; D001894:Borohydrides; D007295:Inositol Phosphates; D010446:Peptide Fragments; C002647:inositol 1-phosphate; D010669:Phenylthiohydantoin; D011732:Pyridoxal Phosphate; C025364:sodium borohydride; D010744:Phosphoric Monoester Hydrolases; C021822:myo-inositol-1 (or 4)-monophosphatase; D008239:Lysine