The butyrylcholinesterase K-variant shows similar cellular protein turnover and quaternary interaction to the wild-type enzyme

Cibby Varkey Altamirano, Cynthia F. Bartels, Oksana Lockridge

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

A recent study has linked the butyrylcholinesterase (BChE) K-variant and the apolipoprotein ε4 isoform to late-onset Alzheimer's disease. These findings have been controversial and have led us to examine the differences between wild-type and K-variant BChE in enzyme activity, protein stability, and quaternary structure. J-variant BChE (E497V/A539T) was also studied because it is associated with the K-variant mutation. The K-variant mutation (A539T) is located in the C-terminal tetramerization domain. Wild-type, K- variant, and J-variant BChE were expressed in Chinese hamster ovary cells and purified. The purified enzymes had similar binding affinity (K(m)) values and catalytic rates for butyrylthiocholine and benzoylcholine. In pulse-chase studies the K-variant, J-variant, and wildtype BChE were degraded rapidly within the cell, with a half-time of ~1.5 h. Less than 5% of the intracellular BChE was exported. The C-terminal peptide containing the K- variant mutation interacted with itself as strongly as did the wild-type peptide in the yeast two-hybrid system. Both K-variant and wild-type BChE assembled into tetramers in the presence of poly-L-proline or the proline- rich attachment domain of the collagen tail. The native K-variant BChE in serum showed the same proportion of tetramers as the native serum wild-type BChE. We conclude that the K-variant BChE is similar to wild-type BChE in enzyme activity, protein turnover, and tetramer formation.

Original languageEnglish (US)
Pages (from-to)869-877
Number of pages9
JournalJournal of Neurochemistry
Volume74
Issue number2
DOIs
StatePublished - Jan 29 2000

Fingerprint

Butyrylcholinesterase
Enzymes
Proteins
Enzyme activity
Mutation
Benzoylcholine
Butyrylthiocholine
Peptides
Two-Hybrid System Techniques
Apolipoproteins
Protein Stability
Cricetulus
Serum
Hybrid systems
Proline
Yeast
Tail
Ovary
Alzheimer Disease
Protein Isoforms

Keywords

  • Butyrylcholinesterase
  • K-variant
  • Late-onset Alzheimer's disease

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

The butyrylcholinesterase K-variant shows similar cellular protein turnover and quaternary interaction to the wild-type enzyme. / Altamirano, Cibby Varkey; Bartels, Cynthia F.; Lockridge, Oksana.

In: Journal of Neurochemistry, Vol. 74, No. 2, 29.01.2000, p. 869-877.

Research output: Contribution to journalArticle

@article{80640a29ebe44ded959da2e88ce65689,
title = "The butyrylcholinesterase K-variant shows similar cellular protein turnover and quaternary interaction to the wild-type enzyme",
abstract = "A recent study has linked the butyrylcholinesterase (BChE) K-variant and the apolipoprotein ε4 isoform to late-onset Alzheimer's disease. These findings have been controversial and have led us to examine the differences between wild-type and K-variant BChE in enzyme activity, protein stability, and quaternary structure. J-variant BChE (E497V/A539T) was also studied because it is associated with the K-variant mutation. The K-variant mutation (A539T) is located in the C-terminal tetramerization domain. Wild-type, K- variant, and J-variant BChE were expressed in Chinese hamster ovary cells and purified. The purified enzymes had similar binding affinity (K(m)) values and catalytic rates for butyrylthiocholine and benzoylcholine. In pulse-chase studies the K-variant, J-variant, and wildtype BChE were degraded rapidly within the cell, with a half-time of ~1.5 h. Less than 5{\%} of the intracellular BChE was exported. The C-terminal peptide containing the K- variant mutation interacted with itself as strongly as did the wild-type peptide in the yeast two-hybrid system. Both K-variant and wild-type BChE assembled into tetramers in the presence of poly-L-proline or the proline- rich attachment domain of the collagen tail. The native K-variant BChE in serum showed the same proportion of tetramers as the native serum wild-type BChE. We conclude that the K-variant BChE is similar to wild-type BChE in enzyme activity, protein turnover, and tetramer formation.",
keywords = "Butyrylcholinesterase, K-variant, Late-onset Alzheimer's disease",
author = "Altamirano, {Cibby Varkey} and Bartels, {Cynthia F.} and Oksana Lockridge",
year = "2000",
month = "1",
day = "29",
doi = "10.1046/j.1471-4159.2000.740869.x",
language = "English (US)",
volume = "74",
pages = "869--877",
journal = "Journal of Neurochemistry",
issn = "0022-3042",
publisher = "Wiley-Blackwell",
number = "2",

}

TY - JOUR

T1 - The butyrylcholinesterase K-variant shows similar cellular protein turnover and quaternary interaction to the wild-type enzyme

AU - Altamirano, Cibby Varkey

AU - Bartels, Cynthia F.

AU - Lockridge, Oksana

PY - 2000/1/29

Y1 - 2000/1/29

N2 - A recent study has linked the butyrylcholinesterase (BChE) K-variant and the apolipoprotein ε4 isoform to late-onset Alzheimer's disease. These findings have been controversial and have led us to examine the differences between wild-type and K-variant BChE in enzyme activity, protein stability, and quaternary structure. J-variant BChE (E497V/A539T) was also studied because it is associated with the K-variant mutation. The K-variant mutation (A539T) is located in the C-terminal tetramerization domain. Wild-type, K- variant, and J-variant BChE were expressed in Chinese hamster ovary cells and purified. The purified enzymes had similar binding affinity (K(m)) values and catalytic rates for butyrylthiocholine and benzoylcholine. In pulse-chase studies the K-variant, J-variant, and wildtype BChE were degraded rapidly within the cell, with a half-time of ~1.5 h. Less than 5% of the intracellular BChE was exported. The C-terminal peptide containing the K- variant mutation interacted with itself as strongly as did the wild-type peptide in the yeast two-hybrid system. Both K-variant and wild-type BChE assembled into tetramers in the presence of poly-L-proline or the proline- rich attachment domain of the collagen tail. The native K-variant BChE in serum showed the same proportion of tetramers as the native serum wild-type BChE. We conclude that the K-variant BChE is similar to wild-type BChE in enzyme activity, protein turnover, and tetramer formation.

AB - A recent study has linked the butyrylcholinesterase (BChE) K-variant and the apolipoprotein ε4 isoform to late-onset Alzheimer's disease. These findings have been controversial and have led us to examine the differences between wild-type and K-variant BChE in enzyme activity, protein stability, and quaternary structure. J-variant BChE (E497V/A539T) was also studied because it is associated with the K-variant mutation. The K-variant mutation (A539T) is located in the C-terminal tetramerization domain. Wild-type, K- variant, and J-variant BChE were expressed in Chinese hamster ovary cells and purified. The purified enzymes had similar binding affinity (K(m)) values and catalytic rates for butyrylthiocholine and benzoylcholine. In pulse-chase studies the K-variant, J-variant, and wildtype BChE were degraded rapidly within the cell, with a half-time of ~1.5 h. Less than 5% of the intracellular BChE was exported. The C-terminal peptide containing the K- variant mutation interacted with itself as strongly as did the wild-type peptide in the yeast two-hybrid system. Both K-variant and wild-type BChE assembled into tetramers in the presence of poly-L-proline or the proline- rich attachment domain of the collagen tail. The native K-variant BChE in serum showed the same proportion of tetramers as the native serum wild-type BChE. We conclude that the K-variant BChE is similar to wild-type BChE in enzyme activity, protein turnover, and tetramer formation.

KW - Butyrylcholinesterase

KW - K-variant

KW - Late-onset Alzheimer's disease

UR - http://www.scopus.com/inward/record.url?scp=0033971911&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033971911&partnerID=8YFLogxK

U2 - 10.1046/j.1471-4159.2000.740869.x

DO - 10.1046/j.1471-4159.2000.740869.x

M3 - Article

C2 - 10646540

AN - SCOPUS:0033971911

VL - 74

SP - 869

EP - 877

JO - Journal of Neurochemistry

JF - Journal of Neurochemistry

SN - 0022-3042

IS - 2

ER -