Isolation of acetylcholinesterase from apoptotic human lung fibroblast cells by antibody affinity chromatography

Qi Huang Jin, Yu Fang Shi, Heng Yi He, Kelvin K.W. Ng, Hua Jiang, Lei Yang, Zi Qing Jiang, Xue Jun Zhang

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Acetylcholinesterase (AChE; EC3.1.1.7) is well known for its role in the hydrolysis of acetylcholine at cholinergic synapses to terminate neurotransmission. In addition to its synaptic presence, AChE has been found to be in non-cholinergic cells such as hematopoietic and osteogenic cells. We have recently reported that AChE is expressed in various cells undergoing apoptosis. To characterize AChE in apoptotic cells and to investigate the role of AChE expression in apoptosis, we devised a method to purify AChE expressed in apoptotic human lung fibroblast cell line HLF. The isolation of this enzyme is mainly based on inhibitor ligand affinity chromatography using immobilized tacrine. However, this method is only effective in isolating active AChE. Here we employed antibody-based chromatography and found that both active and inactive AChE were present in apoptotic HLF cells. Active AChE was predominantly observed in the nuclei of apoptotic cells, while inactive AChE was mainly present in the cytoplasm. Therefore, our method provides an opportunity to investigate further the role of AChE, especially inactive AChE, in apoptosis.

Original languageEnglish (US)
JournalBioTechniques
Volume33
Issue number4 SUPPL.
StatePublished - Oct 1 2002

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Affinity chromatography
Antibody Affinity
Fibroblasts
Acetylcholinesterase
Affinity Chromatography
Cells
Apoptosis
Lung
Antibodies
Tacrine
Chromatography
Cholinergic Agents
Acetylcholine
Hydrolysis
Ligands
Cell Nucleus
Synaptic Transmission
Synapses
Enzymes
Cytoplasm

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Jin, Q. H., Shi, Y. F., He, H. Y., Ng, K. K. W., Jiang, H., Yang, L., ... Zhang, X. J. (2002). Isolation of acetylcholinesterase from apoptotic human lung fibroblast cells by antibody affinity chromatography. BioTechniques, 33(4 SUPPL.).

Isolation of acetylcholinesterase from apoptotic human lung fibroblast cells by antibody affinity chromatography. / Jin, Qi Huang; Shi, Yu Fang; He, Heng Yi; Ng, Kelvin K.W.; Jiang, Hua; Yang, Lei; Jiang, Zi Qing; Zhang, Xue Jun.

In: BioTechniques, Vol. 33, No. 4 SUPPL., 01.10.2002.

Research output: Contribution to journalArticle

Jin, QH, Shi, YF, He, HY, Ng, KKW, Jiang, H, Yang, L, Jiang, ZQ & Zhang, XJ 2002, 'Isolation of acetylcholinesterase from apoptotic human lung fibroblast cells by antibody affinity chromatography', BioTechniques, vol. 33, no. 4 SUPPL..
Jin, Qi Huang ; Shi, Yu Fang ; He, Heng Yi ; Ng, Kelvin K.W. ; Jiang, Hua ; Yang, Lei ; Jiang, Zi Qing ; Zhang, Xue Jun. / Isolation of acetylcholinesterase from apoptotic human lung fibroblast cells by antibody affinity chromatography. In: BioTechniques. 2002 ; Vol. 33, No. 4 SUPPL.
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