Expression of Soluble Ligand- and Antibody-binding Extracellular Domain of Human Muscle Acetylcholine Receptor α Subunit in Yeast Pichia pastoris: ROLE OF GLYCOSYLATION IN α-BUNGAROTOXIN BINDING
Loukia Psaridi-Linardaki,Avgi Mamalaki,Michael Remoundos,Socrates J. Tzartos,Socrates J. Tzartos +4 more
TLDR
Results suggest that the extracellular domain of the human AChR α subunit expressed in P. pastoris has an apparently near native conformation, which makes it suitable for structural studies on the nicotinic acetylcholine receptor and for use as an autoantigen in myasthenia gravis studies.About:
This article is published in Journal of Biological Chemistry.The article was published on 2002-07-26 and is currently open access. It has received 58 citations till now. The article focuses on the topics: Acetylcholine receptor & Pichia pastoris.read more
Citations
More filters
Journal ArticleDOI
Expression of heterologous proteins in Pichia pastoris: a useful experimental tool in protein engineering and production
Rachel Daly,Milton T.W. Hearn +1 more
TL;DR: The purpose of this review is to summarize important developments and features of this expression system and to examine from an experimental perspective the genetic engineering, protein chemical and molecular design considerations that have to be taken into account for the successful expression of the target recombinant protein.
Journal ArticleDOI
Crystal structure of the extracellular domain of nAChR alpha1 bound to alpha-bungarotoxin at 1.94 A resolution.
TL;DR: This structure is the first atomic-resolution view of a nAChR subunit extracellular domain, revealing receptor-specific features such as the main immunogenic region (MIR), the signature Cys-loop and the N-linked carbohydrate chain.
Journal ArticleDOI
Muscle and neuronal nicotinic acetylcholine receptors
Dimitra Kalamida,Konstantinos Poulas,Konstantinos Poulas,Vassiliki Avramopoulou,Efrosini Fostieri,George Lagoumintzis,Konstantinos Lazaridis,Anastasia Sideri,Marios Zouridakis,Marios Zouridakis,Socrates J. Tzartos,Socrates J. Tzartos +11 more
TL;DR: Nicotinic acetylcholine receptors (nAChRs) as mentioned in this paper are integral membrane proteins and prototypic members of the ligand-gated ion-channel superfamily, which has precursors in the prokaryotic world.
Journal ArticleDOI
Antibodies in Myasthenia Gravis and Related Disorders
Angela Vincent,John McConville,Maria Elena Farrugia,John Bowen,Paul Plested,Teresa Tang,Amelia Evoli,Ian Matthews,Gary Sims,Paola Dalton,Leslie Jacobson,Agata Polizzi,Frans Blaes,Bethan Lang,David Beeson,Nick Willcox,John Newsom-Davis,Werner Hoch +17 more
TL;DR: Acetylcholine receptor (AChR) antibodies are present in around 85% of patients with myasthenia gravis as measured by the conventional radioimmunoprecipitation assay, and MuSK antibody positivity appears to be associated with more severe bulbar disease that can be difficult to treat effectively.
Journal ArticleDOI
Crystal structures of free and antagonist-bound states of human α9 nicotinic receptor extracellular domain
Marios Zouridakis,Petros Giastas,Eleftherios Zarkadas,Dafni Chroni-Tzartou,Piotr Bregestovski,Socrates J. Tzartos,Socrates J. Tzartos +6 more
TL;DR: The X-ray crystal structures of the α9 ECD revealed a functionally important β7-β10 strand interaction in α9-containing nAChRs, involving their unique Thr147, a hydration pocket similar to that of mouse α1 ECD and a membrane-facing network coordinated by the invariant Arg210.
References
More filters
Journal ArticleDOI
Heterologous protein expression in the methylotrophic yeast Pichia pastoris
TL;DR: This paper reviews the P. pastoris expression system: how it was developed, how it works, and what proteins have been produced and describes new promoters and auxotrophic marker/host strain combinations which extend the usefulness of the system.
Journal ArticleDOI
Crystal structure of an ACh-binding protein reveals the ligand-binding domain of nicotinic receptors.
Katjuša Brejc,W.J. van Dijk,Remco V. Klaassen,M. Schuurmans,J. van der Oost,J. van der Oost,August B. Smit,Titia K. Sixma +7 more
TL;DR: The crystal structure of molluscan acetylcholine-binding protein (AChBP), a structural and functional homologue of the amino-terminal ligand-binding domain of an nAChR α-subunit, is presented and is relevant for the development of drugs against Alzheimer’s disease and nicotine addiction.
Journal ArticleDOI
Recombinant protein expression in Pichia pastoris.
TL;DR: Major advances in the development of new strains and vectors, improved techniques, and the commercial availability of these tools coupled with a better understanding of the biology of Pichia species have led to this microbe’s value and power in commercial and research labs alike.
Journal ArticleDOI
Toward a structural basis for the function of nicotinic acetylcholine receptors and their cousins
Arthur Karlin,Myles H. Akabas +1 more
TL;DR: The nicotinic acetylcholine (ACh) receptors and the other neurotransmitter-gated ion channels have key roles in fast synaptic transmission throughout the nervous system and new information about the parts of these receptors that are in the front line of function, the neurotransmitter binding sites, the ion-conducting channel, and the gate provides intriguing clues about the mechanisms.
Journal ArticleDOI
Nicotinic acetylcholine receptor at 4.6 A resolution: transverse tunnels in the channel wall.
TL;DR: It is suggested that the extracellular tunnels are access routes to the binding pockets for ACh, and that the cytoplasmic openings serve as filters to exclude anions and other impermeant species from the vicinity of the pore.