Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Intramolecular relationships in cholinesterases revealed by oocyte expression of site-directed and natural variants of human BCHE.
Neville LF
,
Gnatt A
,
Loewenstein Y
,
Seidman S
,
Ehrlich G
,
Soreq H
.
???displayArticle.abstract???
Structure-function relationships of cholinesterases (CHEs) were studied by expressing site-directed and naturally occurring mutants of human butyrylcholinesterase (BCHE) in microinjected Xenopus oocytes. Site-directed mutagenesis of the conserved electronegative Glu441,Ile442,Glu443 domain to Gly441,Ile442,Gln443 drastically reduced the rate of butyrylthiocholine (BTCh) hydrolysis and caused pronounced resistance to dibucaine binding. These findings implicate the charged Glu441,Ile442,Glu443 domain as necessary for a functional CHE catalytic triad as well as for binding quinoline derivatives. Asp70 to Gly substitution characteristic of 'atypical' BCHE, failed to alter its Km towards BTCh or dibucaine binding but reduced hydrolytic activity to 25% of control. Normal hydrolytic activity was restored to Gly70 BCHE by additional His114 or Tyr561 mutations, both of which co-appear with Gly70 in natural BCHE variants, which implies a likely selection advantage for these double BCHE mutants over the single Gly70 BCHE variant. Gly70 BCHE variants also displayed lower binding as compared with Asp70 BCHE to cholinergic drugs, certain choline esters and solanidine. These effects were ameliorated in part by additional mutations or in binding solanidine complexed with sugar residues. These observations indicate that structural interactions exist between N' and C' terminal domains in CHEs which contribute to substrate and inhibitor binding and suggest a crucial involvement of both electrostatic and hydrophobic domains in the build-up of the CHE active center.
Ben Aziz,
Improving poor in vitro transcription from G,C-rich genes.
1990, Pubmed
Ben Aziz,
Improving poor in vitro transcription from G,C-rich genes.
1990,
Pubmed
Berman,
Fluorescent phosphonate labels for serine hydrolases. Kinetic and spectroscopic properties of (7-nitrobenz-2-oxa-1,3-diazole)aminoalkyl methylphosphonofluoridates and their conjugates with acetylcholinesterase molecular forms.
1985,
Pubmed
Brady,
A serine protease triad forms the catalytic centre of a triacylglycerol lipase.
1990,
Pubmed
Brennan,
Hypermutability of CpG dinucleotides in the propeptide-encoding sequence of the human albumin gene.
1990,
Pubmed
Cohen,
1-Bromopinacolone, an active site-directed covalent inhibitor for acetylcholinesterase.
1982,
Pubmed
Gibney,
Mutagenesis of essential functional residues in acetylcholinesterase.
1990,
Pubmed
Gnatt,
Expression of alternatively terminated unusual human butyrylcholinesterase messenger RNA transcripts, mapping to chromosome 3q26-ter, in nervous system tumors.
1990,
Pubmed
HARRIS,
Differential inhibition of the serum cholinesterase phenotypes by solanine and solanidine.
1962,
Pubmed
HODGKIN,
COMPLETE PSEUDOCHOLINESTERASE DEFICIENCY: GENETIC AND IMMUNOLOGIC CHARACTERIZATION.
1965,
Pubmed
Hasan,
Cationic and uncharged substrates and reversible inhibitors in hydrolysis by acetylcholinesterase (EC 3.1.1.7). The trimethyl subsite.
1981,
Pubmed
Hasan,
Hydrolysis by acetylcholinesterase. Apparent molal volumes and trimethyl and methyl subsites.
1980,
Pubmed
Kragh-Hansen,
Binding of warfarin, salicylate, and diazepam to genetic variants of human serum albumin with known mutations.
1990,
Pubmed
Kunkel,
Rapid and efficient site-specific mutagenesis without phenotypic selection.
1987,
Pubmed
La Du,
Proposed nomenclature for human butyrylcholinesterase genetic variants identified by DNA sequencing.
1991,
Pubmed
Lapidot-Lifson,
Coamplification of human acetylcholinesterase and butyrylcholinesterase genes in blood cells: correlation with various leukemias and abnormal megakaryocytopoiesis.
1989,
Pubmed
Layer,
Spatiotemporal relationship of embryonic cholinesterases with cell proliferation in chicken brain and eye.
1987,
Pubmed
Lockridge,
Location of disulfide bonds within the sequence of human serum cholinesterase.
1987,
Pubmed
MacPhee-Quigley,
Profile of the disulfide bonds in acetylcholinesterase.
1986,
Pubmed
Majumdar,
Chemical modification of acetylcholinesterase from eel and basal ganglia: effect on the acetylcholinesterase and aryl acylamidase activities.
1984,
Pubmed
Naveh,
New substrates of acetylcholinesterase.
1981,
Pubmed
Neville,
Anionic site interactions in human butyrylcholinesterase disrupted by two single point mutations.
1990,
Pubmed
,
Xenbase
Neville,
Aspartate-70 to glycine substitution confers resistance to naturally occurring and synthetic anionic-site ligands on in-ovo produced human butyrylcholinesterase.
1990,
Pubmed
,
Xenbase
Nolte,
Effective charge on acetylcholinesterase active sites determined from the ionic strength dependence of association rate constants with cationic ligands.
1980,
Pubmed
Page,
Acetylcholinesterase: inhibition by tetranitromethane and arsenite. Binding of arsenite by tyrosine residues.
1985,
Pubmed
Patinkin,
Manipulations of cholinesterase gene expression modulate murine megakaryocytopoiesis in vitro.
1990,
Pubmed
,
Xenbase
Piela,
Proline-induced constraints in alpha-helices.
1987,
Pubmed
Prody,
Isolation and characterization of full-length cDNA clones coding for cholinesterase from fetal human tissues.
1987,
Pubmed
Russell,
Electrostatic effects on modification of charged groups in the active site cleft of subtilisin by protein engineering.
1987,
Pubmed
Russell,
Rational modification of enzyme catalysis by engineering surface charge.
,
Pubmed
Sali,
Stabilization of protein structure by interaction of alpha-helix dipole with a charged side chain.
1988,
Pubmed
Shinitzky,
Spectral evidence for the presence of tryptophan in the binding site of acetylcholinesterase.
1973,
Pubmed
Soreq,
Expression and tissue-specific assembly of human butyrylcholine esterase in microinjected Xenopus laevis oocytes.
1989,
Pubmed
,
Xenbase
Sussman,
Atomic structure of acetylcholinesterase from Torpedo californica: a prototypic acetylcholine-binding protein.
1991,
Pubmed
Valentino,
Prediction of drug sensitivity in individuals with atypical serum cholinesterase based on in vitro biochemical studies.
1981,
Pubmed
Weise,
Anionic subsites of the acetylcholinesterase from Torpedo californica: affinity labelling with the cationic reagent N,N-dimethyl-2-phenyl-aziridinium.
1990,
Pubmed
Winkler,
Structure of human pancreatic lipase.
1990,
Pubmed
Zakut,
Chorionic villus cDNA library displays expression of butyrylcholinesterase: putative genetic disposition for ecological danger.
1991,
Pubmed
Zakut,
Polymorphism of acetylcholinesterase in discrete regions of the developing human fetal brain.
1985,
Pubmed
,
Xenbase
Zakut,
Acetylcholinesterase and butyrylcholinesterase genes coamplify in primary ovarian carcinomas.
1990,
Pubmed
,
Xenbase
Zhang,
Toward a simplification of the protein folding problem: a stabilizing polyalanine alpha-helix engineered in T4 lysozyme.
1991,
Pubmed
