The C5 Variant of the Butyrylcholinesterase Tetramer Includes a Noncovalently Bound 60 kDa Lamellipodin Fragment

Schopfer, Lawrence M.; Delacour, Hervé; Masson, Patrick; Leroy, Jacqueline; Krejci, Éric; Lockridge, Oksana

doi:10.3390/molecules22071083

Cited by 16 publications

(9 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Physiologically BChE is mainly a tetramer through the interaction of the C-terminal helices of each monomer that is associated in a coiled coil motif around an antiparallel proline rich peptide originating from lamellipodin [ 26 , 27 , 28 ]. Some recombinant BChE productions and X-ray structures have reported the isolation of dimers of BChE and the tetrameric form can be view as a dimer of dimers.…”

Section: Resultsmentioning

confidence: 99%

Bacterial Expression of Human Butyrylcholinesterase as a Tool for Nerve Agent Bioscavengers Development

Brazzolotto¹,

Igert²,

Guillon³

et al. 2017

Molecules

View full text Add to dashboard Cite

Human butyrylcholinesterase is a performant stoichiometric bioscavenger of organophosphorous nerve agents. It is either isolated from outdated plasma or functionally expressed in eukaryotic systems. Here, we report the production of active human butyrylcholinesterase in a prokaryotic system after optimization of the primary sequence through the Protein Repair One Stop Shop process, a structure- and sequence-based algorithm for soluble bacterial expression of difficult eukaryotic proteins. The mutant enzyme was purified to homogeneity. Its kinetic parameters with substrate are similar to the endogenous human butyrylcholinesterase or recombinants produced in eukaryotic systems. The isolated protein was prone to crystallize and its 2.5-Å X-ray structure revealed an active site gorge region identical to that of previously solved structures. The advantages of this alternate expression system, particularly for the generation of butyrylcholinesterase variants with nerve agent hydrolysis activity, are discussed.

show abstract

Section: Resultsmentioning

confidence: 99%

Bacterial Expression of Human Butyrylcholinesterase as a Tool for Nerve Agent Bioscavengers Development

Brazzolotto¹,

Igert²,

Guillon³

et al. 2017

Molecules

View full text Add to dashboard Cite

show abstract

“…In mammals, AChE and butyrylcholinesterase (BChE) are enzymes that hydrolyze ACh and have similar molecular forms ( Massoulié, 2002 ), both containing a catalytic domain and a similar C-terminal tetramerization domain that requires a proline-rich sequence to organize the tetramer ( Figure 1A ). Several peptides containing polyproline sequences were found in AChE and butyrylcholinesterase (BChE) tetramers ( Li et al, 2008 ; Biberoglu et al, 2012 , 2013 ; Schopfer et al, 2017 ), confirmed in the three-dimensional (3D) structure of BChE tetramer purified from the serum ( Leung et al, 2018 ; Boyko et al, 2019 ); and two proteins ColQ and proline-rich membrane anchor (PRiMA) contain a proline-rich sequence that organizes AChE and/or BChE into tetramers. PRiMA is a small transmembrane protein that anchors AChE tetramer on the plasma membrane ( Perrier et al, 2002 ; Dobbertin et al, 2009 ) and ColQ is specific collagen ( Figures 1A , 2 ) that anchors AChE tetramers in the basal lamina ( Feng et al, 1999 ).…”

Section: Organization Of the Nmj: Cholinesterasesmentioning

confidence: 90%

Cholinesterases in Tripartite Neuromuscular Synapse

Петров

Proskurina

Krejci

2021

Front. Mol. Neurosci.

Self Cite

View full text Add to dashboard Cite

The neuromuscular junction (NMJ) is a tripartite synapse in which not only presynaptic and post-synaptic cells participate in synaptic transmission, but also terminal Schwann cells (TSC). Acetylcholine (ACh) is the neurotransmitter that mediates the signal between the motor neuron and the muscle but also between the motor neuron and TSC. ACh action is terminated by acetylcholinesterase (AChE), anchored by collagen Q (ColQ) in the basal lamina of NMJs. AChE is also anchored by a proline-rich membrane anchor (PRiMA) to the surface of the nerve terminal. Butyrylcholinesterase (BChE), a second cholinesterase, is abundant on TSC and anchored by PRiMA to its plasma membrane. Genetic studies in mice have revealed different regulations of synaptic transmission that depend on ACh spillover. One of the strongest is a depression of ACh release that depends on the activation of α7 nicotinic acetylcholine receptors (nAChR). Partial AChE deficiency has been described in many pathologies or during treatment with cholinesterase inhibitors. In addition to changing the activation of muscle nAChR, AChE deficiency results in an ACh spillover that changes TSC signaling. In this mini-review, we will first briefly outline the organization of the NMJ. This will be followed by a look at the role of TSC in synaptic transmission. Finally, we will review the pathological conditions where there is evidence of decreased AChE activity.

show abstract

“…The BChE tetramer incorporates not only short polyproline-rich peptides, but also long protein fragments that contain a polyproline-rich region. An example is the C5 variant of human BChE whose tetrameric structure includes a 60 kDa lamellipodin fragment [24]. The ability of BChE monomers to assemble into stable, long-lived tetramers by binding the polyproline-rich region of a protein, suggests that BChE could serve as a delivery vehicle for any protein that has been engineered to include a polyproline-rich peptide tag.…”

Section: Bche and Ache Scavenge Polyproline Peptides Released From Proteins In The Cytoplasm Nucleus Endoplasmic Reticulum Extracellular mentioning

confidence: 99%

Polyproline-rich Peptides Organize 4 Cholinesterase Subunits into A Tetramer; BChE and AChE Scavenge Polyproline Peptides Released during Metabolic Turnover

Lockridge

Schopfer

2020

Proceedings of the 2nd International Online Conference on Crystals

Self Cite

View full text Add to dashboard Cite

The genes for acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) encode the proteins responsible for enzyme activity. Additional gene products, PRiMA and ColQ, anchor AChE and BChE proteins into membranes. Soluble AChE and BChE tetramers are composed of 4 identical subunits plus one polyproline-rich peptide. Dilution does not release the polyprolinerich peptide from tetramers. However, protein denaturation, for example heating in a boiling water bath, dissociates the polyproline-rich peptide. Using mass spectrometry to sequence peptides released from soluble AChE and BChE tetramers, we find sequences that correspond to proline-rich regions from a variety of proteins. A typical peptide sequence contains 20 consecutive prolines in a 23-residue peptide LPPPPPPPPPPPPPPPPPPPPLP. There is no single, common consensus sequence i.e., no specific gene appears to be responsible for the polyproline-rich peptides found in soluble AChE and BChE tetramers. We propose that during metabolic turnover, protein fragments containing polyproline-rich sequences are scavenged by AChE and BChE dimers, to make stable AChE and BChE tetramers. The 40-residue, alpha-helical C-terminus of AChE or BChE is the tetramerization domain that binds the polyproline-rich peptide. Four parallel alpha helices wrap around a single antiparallel polyproline peptide to lock the tetramer in place. This organization was established by classical X-ray crystallography for isolated C-termini in complex with a proline-rich peptide. The organization was confirmed for intact, tetrameric human BChE using cryoelectron microscopy. When 40 amino acids are deleted from the carboxy terminus, monomeric enzymes are created that retain full enzymatic activity.

show abstract

The C5 Variant of the Butyrylcholinesterase Tetramer Includes a Noncovalently Bound 60 kDa Lamellipodin Fragment

Cited by 16 publications

References 37 publications

Bacterial Expression of Human Butyrylcholinesterase as a Tool for Nerve Agent Bioscavengers Development

Bacterial Expression of Human Butyrylcholinesterase as a Tool for Nerve Agent Bioscavengers Development

Cholinesterases in Tripartite Neuromuscular Synapse

Polyproline-rich Peptides Organize 4 Cholinesterase Subunits into A Tetramer; BChE and AChE Scavenge Polyproline Peptides Released during Metabolic Turnover

Contact Info

Product

Resources

About