The PRiMA-linked Cholinesterase Tetramers Are Assembled from Homodimers

Chen, Vicky P.; Xie, Heidi Qunhui; Chan, Wallace K.B.; Leung, Ka Wing; Chan, Gallant Kar-Lun; Choi, Roy Chi Yan; Bon, Suzanne; Massoulié, Jean; Tsim, Karl Wah Keung

doi:10.1074/jbc.m110.113647

Cited by 22 publications

(17 citation statements)

References 45 publications

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“…4C). Light dimers (no PRiMA), heavy dimers (with PRiMA), and tetramers (with PRiMA) were observed in the G 4 -enriched fractions, as reported previously (21). We obtained identical results in the case of AChE T N296Q/N381Q/N495Q , which suggested the oligomerization of AChE dimer or tetramer was not affected by the glycosylation of AChE T .…”

Section: Resultssupporting

confidence: 77%

“…We have shown that mammalian and chicken AChE T subunits can form mixed AChE T dimers or tetramers, in agreement with the notion that this association depends on the "four-helixbundle" contact between two catalytic domains (21). To investigate whether glycosylated and un-glycosylated AChE T subunits possess the same capacity for dimeric or AChE T N296Q/N381Q/N495Q and PRiMA-HA.…”

Section: Formation Of Interspecies Chicken-human Ache G 4 Hybrids-mentioning

confidence: 96%

“…Neither antibody cross-reacted with the other corresponding antigen (21). Each sample was added with 30 l of protein G-agarose (Roche Applied Science) and incubated for 4 h at 4°C.…”

Section: Methodsmentioning

confidence: 99%

“…The cDNA encoding full-length mouse PRiMA, tagged with an HA epitope (YPY DVP DYA) at the C terminus, was subcloned in a pEF-BOS vector under the control of an EF-1␣ promoter (4,19). The cDNA encoding full-length chicken AChE T was subcloned in a pcDNA3 vector under a CMV promoter (20,21). One day after plating, cultured HZK293T cells were transfected with cDNAs by calcium phosphate precipitation (22).…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, PRiMA is able to recruit human and chicken AChE T subunits into mixed tetramers, even for the un-glycosylated AChE T N296Q/N381Q/N495Q subunits. The formation of a dimer involves a disulfide linkage near the C terminus of the two AChE T subunits (21). To further confirm the formation of interspecies dimers between AChE T N296Q/N381Q/N495Q and cAChE T , we performed non-reducing gel electrophoresis and Western blotting (Fig.…”

Section: Formation Of Interspecies Chicken-human Ache G 4 Hybrids-mentioning

confidence: 99%

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The Assembly of Proline-rich Membrane Anchor (PRiMA)-linked Acetylcholinesterase Enzyme

Chen

Choi

Chan

et al. 2011

Journal of Biological Chemistry

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Acetylcholinesterase (AChE) anchors onto cell membranes by a transmembrane protein PRiMA (proline-rich membrane anchor) as a tetrameric form in vertebrate brain. The assembly of AChE tetramer with PRiMA requires the C-terminal "t-peptide" in AChE catalytic subunit (AChE T ). Although mature AChE is well known N-glycosylated, the role of glycosylation in forming the physiologically active PRiMA-linked AChE tetramer has not been studied. Here, several lines of evidence indicate that the N-linked glycosylation of AChE T plays a major role for acquisition of AChE full enzymatic activity but does not affect its oligomerization. The expression of the AChE T mutant, in which all N-glycosylation sites were deleted, together with PRiMA in HEK293T cells produced a glycan-depleted PRiMAlinked AChE tetramer but with a much higher K m value as compared with the wild type. This glycan-depleted enzyme was assembled in endoplasmic reticulum but was not transported to Golgi apparatus or plasma membrane.

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Section: Resultssupporting

confidence: 77%

Section: Formation Of Interspecies Chicken-human Ache G 4 Hybrids-mentioning

confidence: 96%

“…Neither antibody cross-reacted with the other corresponding antigen (21). Each sample was added with 30 l of protein G-agarose (Roche Applied Science) and incubated for 4 h at 4°C.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Formation Of Interspecies Chicken-human Ache G 4 Hybrids-mentioning

confidence: 99%

See 3 more Smart Citations

The Assembly of Proline-rich Membrane Anchor (PRiMA)-linked Acetylcholinesterase Enzyme

Chen

Choi

Chan

et al. 2011

Journal of Biological Chemistry

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Hypercholesterolemia induces short-term spatial memory impairments in mice: up-regulation of acetylcholinesterase activity as an early and causal event?

et al. 2013

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Epidemiological studies have indicated hypercholesterolemia in midlife as a risk factor for dementia in later life, bringing cholesterol to the forefront of Alzheimer's disease research. Herein, we modeled mild hypercholesterolemia in mice to evaluate biochemical and behavioral alterations linked to hypercholesterolemia. Swiss mice were fed a high fat/cholesterol diet (20 % fat and 1.25 % cholesterol) for an 8-week period (from 12 to 18 weeks old) and were tested on the object location, forced swimming and elevated plus-maze tasks. We also investigated hypercholesterolemia-induced changes on acetylcholinesterase (AChE) activity, oxidative damage, amyloid precursor protein (APP) processing and blood brain barrier (BBB) integrity within the prefrontal cortex and hippocampus. It was found that increased AChE activity within the prefrontal cortex and hippocampus is an early event associated with hypercholesterolemia-induced short-term memory impairments. We observed no signs of antioxidant imbalance and/or oxidative damage or changes in cortical and hippocampal densities of beta-site amyloid precursor protein-cleaving enzyme 1 and aquaporin-4, biomarkers of APP processing and BBB integrity, respectively. In addition, we treated SH-SY5Y human neuroblastoma cells with low-density lipoprotein (LDL) cholesterol in an attempt to manipulate cell cholesterol content. Notably, LDL cholesterol increased in a dose-dependent manner the activity of AChE in SH-SY5Y cells. The present findings provide new evidence that increased AChE activity within the prefrontal cortex and hippocampus is an early event associated with hypercholesterolemia-induced cognitive impairments.

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Characterizations of Cholinesterases in Golden Apple Snail (Pomacea canaliculata)

et al. 2013

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Cholinesterases (ChEs) have been identified in vertebrates and invertebrates. Inhibition of ChE activity in invertebrates, such as bivalve molluscs, has been used to evaluate the exposure of organophosphates, carbamate pesticides, and heavy metals in the marine system. The golden apple snail (Pomacea canaliculata) is considered as one of the worst invasive alien species harmful to rice and other crops. The ChE(s) in this animal, which has been found recently, but poorly characterized thus far, could serve as biomarker(s) for environmental surveillance as well as a potential target for the pest control. In this study, the tissue distribution, substrate preference, sensitivity to ChE inhibitors, and molecular species of ChEs in P. canaliculata were investigated. It was found that the activities of both AChE and BChE were present in all test tissues. The intestine had the most abundant ChE activities. Both enzymes had fair activities in the head, kidney, and gills. The BChE activity was more sensitive to tetra-isopropylpyrophosphoramide (iso-OMPA) than the AChE. Only one BChE molecular species, 5.8S, was found in the intestine and head, whereas two AChE species, 5.8S and 11.6S, were found there. We propose that intestine ChEs of this snail may be potential biomarkers for manipulating pollutions.

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The PRiMA-linked Cholinesterase Tetramers Are Assembled from Homodimers

Cited by 22 publications

References 45 publications

The Assembly of Proline-rich Membrane Anchor (PRiMA)-linked Acetylcholinesterase Enzyme

The Assembly of Proline-rich Membrane Anchor (PRiMA)-linked Acetylcholinesterase Enzyme

Hypercholesterolemia induces short-term spatial memory impairments in mice: up-regulation of acetylcholinesterase activity as an early and causal event?

Characterizations of Cholinesterases in Golden Apple Snail (Pomacea canaliculata)

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