Characterization of an Autoreduction Pathway for the [Fe4S4]3+Cluster of MutantChromatium vinosumHigh-Potential Iron Proteins. Site-Directed Mutagenesis Studies To Probe the Role of Phenylalanine 66 in Defining the Stability of the [Fe4S4] Center Provide Evidence for Oxidative Degradation via a [Fe3S4] Cluster†

Biochemistry ◽  
1996 ◽  
Vol 35 (46) ◽  
pp. 14544-14552 ◽  
Author(s):  
Shumin Bian ◽  
C. F. Hemann ◽  
Russ Hille ◽  
J. A. Cowan
Keyword(s):  
Oxidative Degradation ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Iron Proteins ◽  
The Stability

Site-directed mutagenesis and homology modeling indicate an important role of cysteine 439 in the stability of betaine aldehyde dehydrogenase from Pseudomonas aeruginosa

Biochimie ◽  
2005 ◽  
Vol 87 (12) ◽  
pp. 1056-1064 ◽  
Author(s):  
Lilian González-Segura ◽  
Roberto Velasco-García ◽  
Enrique Rudiño-Piñera ◽  
Carlos Mújica-Jiménez ◽  
Rosario A. Muñoz-Clares
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Homology Modeling ◽  
Aldehyde Dehydrogenase ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Betaine Aldehyde Dehydrogenase ◽  
Betaine Aldehyde ◽  
The Stability

scholarly journals Kinetic Studies on CphA Mutants Reveal the Role of the P158-P172 Loop in Activity versus Carbapenems

2016 ◽  
Vol 60 (5) ◽  
pp. 3123-3126 ◽  
Author(s):  
Carlo Bottoni ◽  
Mariagrazia Perilli ◽  
Francesca Marcoccia ◽  
Alessandra Piccirilli ◽  
Cristina Pellegrini ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Catalytic Efficiency ◽  
Kinetic Studies ◽  
Zinc Ion ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Wild Type ◽  
The Stability

ABSTRACTSite-directed mutagenesis of CphA indicated that prolines in the P158-P172 loop are essential for the stability and the catalytic activity of subclass B2 metallo-β-lactamases against carbapenems. The sequential substitution of proline led to a decrease of the catalytic efficiency of the variant compared to the wild-type (WT) enzyme but also to a higher affinity for the binding of the second zinc ion.

scholarly journals The Role of the Disulfide Bridge in the Stability and Structural Integrity of Ovalbumin Evaluated by Site-Directed Mutagenesis

2011 ◽  
Vol 75 (3) ◽  
pp. 544-549 ◽  
Author(s):  
Takayuki ISHIMARU ◽  
Kazunari ITO ◽  
Miho TANAKA ◽  
Syunpei TANAKA ◽  
Naotoshi MATSUDOMI
Keyword(s):  
Structural Integrity ◽  
Disulfide Bridge ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
The Stability

Role of tyrosine-80 in the stability of kanamycin nucleotidyltransferase analyzed by site-directed mutagenesis

1988 ◽  
Vol 171 (3) ◽  
pp. 715-720 ◽  
Author(s):  
Masazumi MATSUMURA ◽  
Saburo YAHANDA ◽  
Shigeyoshi YASUMURA ◽  
Katsuhide YUTANI ◽  
Shuichi AIBA
Keyword(s):  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
The Stability

Characterization of Recombinant von Willebrand Factors Mutated on Cysteine 509 or 695

1996 ◽  
Vol 76 (03) ◽  
pp. 453-459 ◽  
Author(s):  
Virginie Siguret ◽  
Anne-Sophie Ribba ◽  
Olivier Christophe ◽  
Ghislaine Chérel ◽  
Bernadette Obert ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Mutant Proteins ◽  
Von Willebrand ◽  
High Molecular Weight Multimers

SummaryThe interacting domain of vWF with platelet GPIb has been shown to overlap the large A1 loop formed by the intra-chain disulfide bond linking Cys 509 to Cys 695. In order to further investigate the role of the conformation of this region, we have expressed in COS-7 cells three mutated full-length recombinant vWFs (rvWFs) in which the substitutions Cys509Gly, Cys509Arg or Cys695Gly have been introduced by site-directed mutagenesis. SDS-agarose gel electrophoresis demonstrated an impaired multimerization of the mutants with undetectable high molecular weight multimers and a decrease of the relative amounts of the intermediate sized multimers. Binding analysis showed that rvWFC509G and rvWFC509R did not interact with botrocetin but spontaneously interacted with GPIb; the latter binding remained unchanged in the presence of ristocetin. This indicates that the substitution of Cys509 by Gly or Arg creates a conformation of vWF that increases its binding to GPIb. In contrast, rvWFC695G which did not react with botrocetin was also unable to interact with GPIb even in the presence of ristocetin, indicating that sequences interacting with GPIb are masked and/or disrupted. In conclusion, the substitution of each of the Cys509 and 695 results in mutant proteins which may be “locked” into active or inactive conformations in regard to the binding to platelet GPIb receptor.

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