Location of functional regions of acetylcholine receptor α-subunit by site-directed mutagenesis

Nature ◽  
1985 ◽  
Vol 313 (6001) ◽  
pp. 364-369 ◽  
Author(s):  
Masayoshi Mishina ◽  
Takamasa Tobimatsu ◽  
Keiji Imoto ◽  
Ken-ichi Tanaka ◽  
Yoshihiko Fujita ◽  
...  
Keyword(s):  
Acetylcholine Receptor ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Α Subunit ◽  
Functional Regions

scholarly journals Site-directed mutagenesis of two conserved charged amino acids in the N-terminal region of α subunit ofE. coli-F0F1

FEBS Letters ◽  
1996 ◽  
Vol 382 (1-2) ◽  
pp. 171-174 ◽  
Author(s):  
Barbara Hase ◽  
Sabine Werner-Grüne ◽  
Gabriele Deckers-Hebestreit ◽  
Heinrich Strotmann
Keyword(s):  
Amino Acids ◽  
Terminal Region ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Α Subunit ◽  
Charged Amino Acids

scholarly journals The functional role of the αM4 transmembrane helix in the muscle nicotinic acetylcholine receptor probed through mutagenesis and coevolutionary analyses

2020 ◽  
Vol 295 (32) ◽  
pp. 11056-11067 ◽  
Author(s):  
Mackenzie J. Thompson ◽  
Jaimee A. Domville ◽  
John E. Baenziger
Keyword(s):  
Acetylcholine Receptor ◽  
Nicotinic Acetylcholine Receptor ◽  
Site Directed Mutagenesis ◽  
Induced Response ◽  
Independent Manner ◽  
Nicotinic Acetylcholine ◽  
Α Subunit ◽  
Lipid Sensing ◽  
Insight Into

The activity of the muscle-type Torpedo nicotinic acetylcholine receptor (nAChR) is highly sensitive to lipids, but the underlying mechanisms remain poorly understood. The nAChR transmembrane α-helix, M4, is positioned at the perimeter of each subunit in direct contact with lipids and likely plays a central role in lipid sensing. To gain insight into the mechanisms underlying nAChR lipid sensing, we used homology modeling, coevolutionary analyses, site-directed mutagenesis, and electrophysiology to examine the role of the α-subunit M4 (αM4) in the function of the adult muscle nAChR. Ala substitutions for most αM4 residues, including those in clusters of polar residues at both the N and C termini, and deletion of up to 11 C-terminal residues had little impact on the agonist-induced response. Even Ala substitutions for coevolved pairs of residues at the interface between αM4 and the adjacent helices, αM1 and αM3, had little effect, although some impaired nAChR expression. On the other hand, Ala substitutions for Thr422 and Arg429 caused relatively large losses of function, suggesting functional roles for these specific residues. Ala substitutions for aromatic residues at the αM4-αM1/αM3 interface generally led to gains of function, as previously reported for the prokaryotic homolog, the Erwinia chrysanthemi ligand-gated ion channel (ELIC). The functional effects of individual Ala substitutions in αM4 were found to be additive, although not in a completely independent manner. Our results provide insight into the structural features of αM4 that are important. They also suggest how lipid-dependent changes in αM4 structure ultimately modify nAChR function.

Introduction of DPR, an Enterostatin Fragment Peptide, into Soybean β-Conglycinin α′ Subunit by Site-directed Mutagenesis

2004 ◽  
Vol 68 (1) ◽  
pp. 253-256 ◽  
Author(s):  
Yasuyuki TAKENAKA ◽  
Naomi DOYAMA ◽  
Nobuyuki MARUYAMA ◽  
Shigeru UTSUMI ◽  
Masaaki YOSHIKAWA
Keyword(s):  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Α Subunit

Site-directed mutagenesis of the α subunit of tryptophan synthase from salmonella typhimurium

1988 ◽  
Vol 151 (2) ◽  
pp. 672-678 ◽  
Author(s):  
Syed Ashrafuddin Ahmed ◽  
Haruhiko Kawasaki ◽  
Ronald Bauerle ◽  
Hatsué Morita ◽  
Edith Wilson Miles
Keyword(s):  
Salmonella Typhimurium ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Tryptophan Synthase ◽  
Α Subunit

Stabilization in microsomal membranes of the fifth transmembrane segment of the Na+,K+-ATPase α subunit with proline to leucine mutation

1993 ◽  
Vol 71 (7-8) ◽  
pp. 410-415 ◽  
Author(s):  
Haruo Homareda ◽  
Kiyoshi Kawakami ◽  
Kei Nagano ◽  
Hideo Matsui
Keyword(s):  
Insertion Site ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Membrane Insertion ◽  
Transmembrane Segment ◽  
Α Subunit ◽  
Microsomal Membranes

We have reported that the fifth transmembrane segment of the Na+,K+-ATPase α subunit could not be inserted into microsomal membranes when it was synthesized from the truncated α1 cDNA. In this study, three proline residues in the segment were changed into leucine residues by site-directed mutagenesis. Replacement of all three proline residues by leucine residues allowed the insertion of this segment and its translocation. This result confirms our previous finding and demonstrates that three proline residues in the fifth transmembrane segment prevent insertion of the segment into the membrane.Key words: Na+,K+-ATPase, transmembrane segment, membrane insertion, site-directed mutagenesis.

Sign in / Sign up

Export Citation Format

Share Document