PMID- 9693127
OWN - NLM
STAT- MEDLINE
DCOM- 19981008
LR  - 20190501
IS  - 0264-6021 (Print)
IS  - 1470-8728 (Electronic)
IS  - 0264-6021 (Linking)
VI  - 334 ( Pt 1)
IP  - Pt 1
DP  - 1998 Aug 15
TI  - Bovine acetylcholinesterase: cloning, expression and characterization.
PG  - 251-9
AB  - The bovine acetylcholinesterase (BoAChE) gene was cloned from genomic DNA and its 
      structure was determined. Five exons coding for the AChE T-subunit and the 
      alternative H-subunit were identified and their organization suggests high 
      conservation of structure in mammalian AChE genes. The deduced amino acid 
      sequence of the bovine T-subunit is highly similar to the human sequence, showing 
      differences at 34 positions only. However, the cloned BoAChE sequence differs 
      from the published amino acid sequence of AChE isolated from fetal bovine serum 
      (FBS) by: (1) 13 amino acids, 12 of which are conserved between BoAChE and human 
      AChE, and (2) the presence of four rather than five potential N-glycosylation 
      sites. The full coding sequence of the mature BoAChE T-subunit was expressed in 
      human embryonal kidney 293 cells (HEK-293). The catalytic properties of 
      recombinant BoAChE and its reactivity towards various inhibitors were similar to 
      those of the native bovine enzyme. Soluble recombinant BoAChE is composed of 
      monomers, dimers and tetramers, yet in contrast to FBS-AChE, tetramer formation 
      is not efficient. Comparative SDS/PAGE analysis reveals that all four potential 
      N-glycosylation sites identified by DNA sequencing appear to be utilized, and 
      that recombinant BoAChE comigrates with FBS-AChE. A major difference between the 
      recombinant enzyme and the native enzyme was observed when clearance from 
      circulation was examined. The HEK-293-derived enzyme was cleared from the 
      circulation at a much faster rate than FBS-AChE. This difference in behaviour, 
      together with previous studies on the effect of post-translation modification on 
      human AChE clearance [Kronman, Velan, Marcus, Ordentlich, Reuveny and Shafferman 
      (1995) Biochem. J. 311, 959-967] suggests that cell-dependent glycosylation plays 
      a key role in AChE circulatory residence.
FAU - Mendelson, I
AU  - Mendelson I
AD  - Department of Biochemistry, Israel Institute for Biological Research, Ness-Ziona, 
      70450, Israel.
FAU - Kronman, C
AU  - Kronman C
FAU - Ariel, N
AU  - Ariel N
FAU - Shafferman, A
AU  - Shafferman A
FAU - Velan, B
AU  - Velan B
LA  - eng
SI  - GENBANK/AF061813
SI  - GENBANK/AF061814
SI  - GENBANK/AF061815
SI  - GENBANK/AF061816
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (Macromolecular Substances)
RN  - 0 (Recombinant Proteins)
RN  - EC 3.1.1.7 (Acetylcholinesterase)
SB  - IM
MH  - Acetylcholinesterase/biosynthesis/blood/*chemistry/*genetics
MH  - Amino Acid Sequence
MH  - Animals
MH  - Cattle
MH  - Cell Line
MH  - Cloning, Molecular
MH  - Dimerization
MH  - Evolution, Molecular
MH  - Fetal Blood/enzymology
MH  - Glycosylation
MH  - Humans
MH  - Kidney
MH  - Macromolecular Substances
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Protein Conformation
MH  - Recombinant Proteins/biosynthesis/chemistry
MH  - Sequence Alignment
MH  - Sequence Homology, Amino Acid
MH  - Transfection
PMC - PMC1219686
EDAT- 1998/08/07 00:00
MHDA- 1998/08/07 00:01
PMCR- 1999/02/15
CRDT- 1998/08/07 00:00
PHST- 1998/08/07 00:00 [pubmed]
PHST- 1998/08/07 00:01 [medline]
PHST- 1998/08/07 00:00 [entrez]
PHST- 1999/02/15 00:00 [pmc-release]
AID - 10.1042/bj3340251 [doi]
PST - ppublish
SO  - Biochem J. 1998 Aug 15;334 ( Pt 1)(Pt 1):251-9. doi: 10.1042/bj3340251.