Identification of Functionally Important Amino Acid Residues within the C2-Domain of Human Factor V Using Alanine-Scanning Mutagenesis†

Biochemistry ◽  
2000 ◽  
Vol 39 (8) ◽  
pp. 1951-1958 ◽  
Author(s):  
Suhng Wook Kim ◽  
Mary Ann Quinn-Allen ◽  
J. Terese Camp ◽  
Sandra Macedo-Ribeiro ◽  
Pablo Fuentes-Prior ◽  
...  
Keyword(s):  
Amino Acid ◽  
Human Factor ◽  
Factor V ◽  
C2 Domain ◽  
Amino Acid Residues ◽  
Alanine Scanning Mutagenesis ◽  
Alanine Scanning ◽  
Important Amino Acid

scholarly journals Alanine scanning of dinitroaniline/phosphorothioamidate site of α-tubulin in plasmodium species distributed in India

2020 ◽  
Vol 26 ◽  
pp. 293-297
Author(s):  
O. M. Demchuk ◽  
P. A. Karpov ◽  
A. V. Rayevsky ◽  
S. P. Ozheredov ◽  
S. I. Spivak ◽  
...  
Keyword(s):  
Protein Structure ◽  
Amino Acid ◽  
Specific Binding ◽  
Plasmodium Species ◽  
Protein Docking ◽  
Amino Acid Residues ◽  
Alanine Scanning Mutagenesis ◽  
Alanine Scanning ◽  
Protein Structure Optimization ◽  
Dynamics Method

Aim. Identification of amino acid residues participating in specific binding of dinitroaniline and phosphorothioamidate compounds with α-tubulin in Plasmodium falciparum. Methods. Protein structure modelling, protein structure optimization using molecular dynamics method, ligand-protein docking, alanine scanning mutagenesis. Results. Molecular docking of canonical compounds and alanine scanning mutagenesis, indicate two key (Arg2, Val250) and one minor (Glu3) residues involved in binding of both - dinitroaniline and phosphorothioamidate compounds. At the same time, it was revealed two minor residues (Asp251, Glu254) interacting only with some members of dinitroaniline grope. Conclusions. It was identified amino acid residues predetermining existence of joint site and similar interaction of α-tubulin with dinitroani-line and phosphorothioamidate compounds in P. falciparum. Keywords: malaria, Plasmodium, α-tubulin, molecular interaction, dinitroanilines compounds, phosphorothioamidate compounds, alanine scanning mutagenesis.

Important Amino Acid Residues that Confer CYP2C19 Selective Activity to CYP2C9

2008 ◽  
Vol 144 (3) ◽  
pp. 323-333 ◽  
Author(s):  
Y. Wada ◽  
M. Mitsuda ◽  
Y. Ishihara ◽  
M. Watanabe ◽  
M. Iwasaki ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Residues ◽  
Important Amino Acid ◽  
Selective Activity

Identification of Important Amino Acid Residues of the Na + -Ca 2+ Exchanger Inhibitory Peptide, XIP

1997 ◽  
Vol 156 (2) ◽  
pp. 149-156 ◽  
Author(s):  
Z. He ◽  
N. Petesch ◽  
K.-P. Voges ◽  
W. Röben ◽  
K.D. Philipson
Keyword(s):  
Amino Acid ◽  
Amino Acid Residues ◽  
Inhibitory Peptide ◽  
Important Amino Acid

The factor V C1 domain is involved in membrane binding: identification of functionally important amino acid residues within the C1 domain of factor V using alanine scanning mutagenesis

2004 ◽  
Vol 91 (01) ◽  
pp. 16-27 ◽  
Author(s):  
Mary Quinn-Allen ◽  
Sandra Macedo-Ribeiro ◽  
Pablo Fuentes-Prior ◽  
Wolfram Bode ◽  
Mahasen Saleh ◽  
...  
Keyword(s):  
Molecular Model ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Aromatic Amino Acids ◽  
Phospholipid Vesicles ◽  
Factor V ◽  
Alanine Scanning Mutagenesis ◽  
Important Amino Acid ◽  
C1 Domain ◽  
Prothrombinase Activity

SummaryThe contribution of the factor Va C1 domain (fVa-C1) to assembly of the prothrombinase complex has not been previously investigated. The homologous fVa-C2 domain contains a binding site for phosphatidylserine (PS) that includes the indole moieties of Trp2063/Trp2064 at the apex of spike-1. In order to investigate the structure and function of fVa-C1 a molecular model was constructed based on the structure of fVa-C2. The aromatic and hydrophobic side chains of Tyr1956/Leu1957 in fVaC1 are located at the predicted apex of spike-3. Exposed charged and hydrophobic residues in fVa-C1 were changed to alanine in clusters of 1-3 mutations per construct. The resultant 20 mutants were expressed in COS cells and screened for binding to immobilized PS and prothrombinase activity on phospholipid vesicles containing either 25% or 5% PS. Two mutants, (Y1956,L1957)A, and (R2023,R2027)A showed both decreased binding to immobilized PS and a selective decrease in prothrombinase activity on membranes containing 5% PS. The interaction of purified (Y1956,L1957)A with phospholipid vesicles was studied using fluorescence resonance energy transfer and prothrombinase assays. The affinity of (Y1956,L1957)A binding to 25% PS membranes was reduced 12-fold compared to rHFVa. Prothrombin activation in the presence of (Y1956,L1957)A was markedly impaired on phospholipid vesicles containing 10% or less PS. We conclude that solvent exposed hydrophobic and aromatic amino acids in both fVa-C1 and fVa-C2 contribute to the interaction of factor V with PS membranes.

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