Probing the inhibitor-binding site of aldose reductase with site-directed mutagenesis

1998 ◽  
Vol 256 (2) ◽  
pp. 310-316 ◽  
Author(s):  
Thomas C. Hohman ◽  
Ossama El-Kabbani ◽  
Michael S. Malamas ◽  
Kehdih Lai ◽  
Tatiana Putilina ◽  
...  
Keyword(s):  
Binding Site ◽  
Aldose Reductase ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Inhibitor Binding

scholarly journals Involvement of cysteine residues in catalysis and inhibition of human aldose reductase. Site-directed mutagenesis of Cys-80, -298, and -303.

1992 ◽  
Vol 267 (34) ◽  
pp. 24833-24840 ◽  
Author(s):  
J.M. Petrash ◽  
T.M. Harter ◽  
C.S. Devine ◽  
P.O. Olins ◽  
A Bhatnagar ◽  
...  
Keyword(s):  
Aldose Reductase ◽  
Site Directed Mutagenesis ◽  
Cysteine Residues ◽  
Directed Mutagenesis

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 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.

Characterization of inhibitor binding to human kinesin spindle protein by site-directed mutagenesis

2009 ◽  
Vol 484 (1) ◽  
pp. 1-7 ◽  
Author(s):  
C. Gary Marshall ◽  
Maricel Torrent ◽  
Olusegun Williams ◽  
Kelly A. Hamilton ◽  
Carolyn A. Buser
Keyword(s):  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Inhibitor Binding ◽  
Kinesin Spindle Protein
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