Identification of the Residues Involved in Stabilization of the Semiquinone Radical in the High-Affinity Ubiquinone Binding Site in Cytochromebo3fromEscherichia coliby Site-Directed Mutagenesis and EPR Spectroscopy†

Biochemistry ◽  
2002 ◽  
Vol 41 (34) ◽  
pp. 10675-10679 ◽  
Author(s):  
Petra Hellwig ◽  
Takahiro Yano ◽  
Tomoko Ohnishi ◽  
Robert B. Gennis
Keyword(s):  
Binding Site ◽  
Epr Spectroscopy ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Semiquinone Radical ◽  
High Affinity ◽  
Ubiquinone Binding

scholarly journals Mapping the BKCa Channel's “Ca2+ Bowl”

2004 ◽  
Vol 123 (5) ◽  
pp. 475-489 ◽  
Author(s):  
Lin Bao ◽  
Christina Kaldany ◽  
Ericka C. Holmstrand ◽  
Daniel H. Cox
Keyword(s):  
Fusion Protein ◽  
Binding Site ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Membrane Domain ◽  
Intracellular Domain ◽  
High Affinity ◽  
Acidic Residues ◽  
The Relationship

There is controversy over whether Ca2+ binds to the BKCa channel's intracellular domain or its integral-membrane domain and over whether or not mutations that reduce the channel's Ca2+ sensitivity act at the point of Ca2+ coordination. One region in the intracellular domain that has been implicated in Ca2+ sensing is the “Ca2+ bowl”. This region contains many acidic residues, and large Ca2+-bowl mutations eliminate Ca2+ sensing through what appears to be one type of high-affinity Ca2+-binding site. Here, through site-directed mutagenesis we have mapped the residues in the Ca2+ bowl that are most important for Ca2+ sensing. We find acidic residues, D898 and D900, to be essential, and we find them essential as well for Ca2+ binding to a fusion protein that contains a portion of the BKCa channel's intracellular domain. Thus, much of our data supports the conclusion that Ca2+ binds to the BKCa channel's intracellular domain, and they define the Ca2+ bowl's essential Ca2+-sensing motif. Overall, however, we have found that the relationship between mutations that disrupt Ca2+ sensing and those that disrupt Ca2+ binding is not as strong as we had expected, a result that raises the possibility that, when examined by gel-overlay, the Ca2+ bowl may be in a nonnative conformation.

scholarly journals The high affinity choline transporter: topology and choline binding site as studied by site‐directed mutagenesis

2008 ◽  
Vol 22 (S1) ◽  
Author(s):  
Kousuke Urashima ◽  
Yuji Kubo ◽  
Shizue Nishikawa ◽  
Toru Nakamura ◽  
Kengo Hayashi ◽  
...  
Keyword(s):  
Binding Site ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Choline Transporter ◽  
High Affinity ◽  
High Affinity Choline Transporter

Probing the Interaction between a High-Affinity Single-Chain Fv and a Pyrimidine (6-4) Pyrimidone Photodimer by Site-Directed Mutagenesis†

Biochemistry ◽  
1999 ◽  
Vol 38 (2) ◽  
pp. 532-539 ◽  
Author(s):  
Hiroyuki Kobayashi ◽  
Hiroshi Morioka ◽  
Kunihiro Tobisawa ◽  
Takuya Torizawa ◽  
Koichi Kato ◽  
...  
Keyword(s):  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Single Chain ◽  
High Affinity ◽  
Single Chain Fv

Protein-ligand interactions in the lysine-binding site of plasminogen kringle 4 are different in crystal and solution. Electrostatic interactions studied by site-directed mutagenesis exclude Lys35 as an important acceptor in solution

Biochemistry ◽  
1993 ◽  
Vol 32 (48) ◽  
pp. 13019-13025 ◽  
Author(s):  
Peter Reinholt Nielsen ◽  
Katja Einer-Jensen ◽  
Thor L. Holtet ◽  
Bente Damm Andersen ◽  
Flemming M. Poulsen ◽  
...  
Keyword(s):  
Binding Site ◽  
Electrostatic Interactions ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Protein Ligand Interactions ◽  
Ligand Interactions

scholarly journals A putative glutathione-binding site in T4 glutaredoxin investigated by site-directed mutagenesis

1991 ◽  
Vol 266 (24) ◽  
pp. 16105-16112
Author(s):  
M. Nikkola ◽  
F.K. Gleason ◽  
M. Saarinen ◽  
T. Joelson ◽  
O. Björnberg ◽  
...  
Keyword(s):  
Binding Site ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Glutathione Binding

scholarly journals Use of Photoaffinity Labeling and Site-directed Mutagenesis for Identification of the Key Residue Responsible for Extraordinarily High Affinity Binding of UCN-01 in Human α1-Acid Glycoprotein

2004 ◽  
Vol 280 (2) ◽  
pp. 1384-1391 ◽  
Author(s):  
Masaaki Katsuki ◽  
Victor Tuan Giam Chuang ◽  
Koji Nishi ◽  
Kohichi Kawahara ◽  
Hitoshi Nakayama ◽  
...  
Keyword(s):  
Photoaffinity Labeling ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Affinity Binding ◽  
High Affinity Binding ◽  
Acid Glycoprotein ◽  
High Affinity
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