Identification of the structural mutation responsible for the dibucaine-resistant (atypical) variant form of human serum cholinesterase

M. C. McGuire, C. P. Nogueira, C. F. Bartels, H. Lightstone, A. Hajra, A. F.L. Van der Spek, O. Lockridge, B. N. La Du

Research output: Contribution to journalArticle

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Abstract

A point mutation in the gene for human serum cholinesterase was identified that changes Asp-70 to Gly in the atypical form of serum cholinesterase. The mutation in nucleotide 209, which changes codon 70 from GAT to GGT, was found by sequencing a genomic clone and sequencing selected regions of DNA amplified by the polymerase chain reaction. The entire coding sequences for usual and atypical cholinesterases were compared, and no other consistent base differences were found. A polymorphic site near the C terminus of the coded region was detected, but neither allele at this locus segregated consistently with the atypical trait. The nucleotide-209 mutation was detected in all five atypical cholinesterase families examined. There was complete concordance between this mutation and serum cholinesterase phenotypes for all 14 heterozygous and 6 homozygous atypical subjects tested. The mutation causes the loss of a Sau3A1 restriction site; the resulting DNA fragment length polymorphism was verified by electrophoresis of 32P-labeled DNA restriction fragments from usual and atypical subjects. Dot-blot hybridization analysis with a 19-mer allele-specific probe to the DNA amplified by the polymerase chain reaction distinguished between the usual and atypical genotypes. We conclude that the Asp-70→Gly mutation (acidic to neutral amino acid substitution) accounts for reduced affinity of atypical cholinesterase for choline esters and that Asp-70 must be an important component of the anionic site. Heterogeneity in atypical alleles may exist, but the Asp-70 point mutation may represent an appreciable portion of the atypical gene pool.

Original languageEnglish (US)
Pages (from-to)953-957
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume86
Issue number3
DOIs
StatePublished - Jan 1 1989

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Dibucaine
Cholinesterases
Mutation
Serum
Alleles
DNA-Directed DNA Polymerase
Point Mutation
Nucleotides
Neutral Amino Acids
Gene Pool
Polymerase Chain Reaction
DNA
Amino Acid Substitution
Choline
Codon
Electrophoresis
Esters
Clone Cells
Genotype
Phenotype

ASJC Scopus subject areas

  • General

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Identification of the structural mutation responsible for the dibucaine-resistant (atypical) variant form of human serum cholinesterase. / McGuire, M. C.; Nogueira, C. P.; Bartels, C. F.; Lightstone, H.; Hajra, A.; Van der Spek, A. F.L.; Lockridge, O.; La Du, B. N.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 86, No. 3, 01.01.1989, p. 953-957.

Research output: Contribution to journalArticle

McGuire, M. C. ; Nogueira, C. P. ; Bartels, C. F. ; Lightstone, H. ; Hajra, A. ; Van der Spek, A. F.L. ; Lockridge, O. ; La Du, B. N. / Identification of the structural mutation responsible for the dibucaine-resistant (atypical) variant form of human serum cholinesterase. In: Proceedings of the National Academy of Sciences of the United States of America. 1989 ; Vol. 86, No. 3. pp. 953-957.
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abstract = "A point mutation in the gene for human serum cholinesterase was identified that changes Asp-70 to Gly in the atypical form of serum cholinesterase. The mutation in nucleotide 209, which changes codon 70 from GAT to GGT, was found by sequencing a genomic clone and sequencing selected regions of DNA amplified by the polymerase chain reaction. The entire coding sequences for usual and atypical cholinesterases were compared, and no other consistent base differences were found. A polymorphic site near the C terminus of the coded region was detected, but neither allele at this locus segregated consistently with the atypical trait. The nucleotide-209 mutation was detected in all five atypical cholinesterase families examined. There was complete concordance between this mutation and serum cholinesterase phenotypes for all 14 heterozygous and 6 homozygous atypical subjects tested. The mutation causes the loss of a Sau3A1 restriction site; the resulting DNA fragment length polymorphism was verified by electrophoresis of 32P-labeled DNA restriction fragments from usual and atypical subjects. Dot-blot hybridization analysis with a 19-mer allele-specific probe to the DNA amplified by the polymerase chain reaction distinguished between the usual and atypical genotypes. We conclude that the Asp-70→Gly mutation (acidic to neutral amino acid substitution) accounts for reduced affinity of atypical cholinesterase for choline esters and that Asp-70 must be an important component of the anionic site. Heterogeneity in atypical alleles may exist, but the Asp-70 point mutation may represent an appreciable portion of the atypical gene pool.",
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