Proton NMR (500 MHz) identification of aromatic residues of gene 32 protein involved in DNA binding by use of protein containing perdeuterated aromatic residues and by site-directed mutagenesis

Biochemistry ◽  
1986 ◽  
Vol 25 (12) ◽  
pp. 3666-3672 ◽  
Author(s):  
Richard V. Prigodich ◽  
Yousif Shamoo ◽  
Kenneth R. Williams ◽  
John W. Chase ◽  
William H. Konigsberg ◽  
...  
Keyword(s):  
Dna Binding ◽  
Proton Nmr ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Aromatic Residues ◽  
Gene 32

scholarly journals DNA‐binding studies of FOXO1 through site‐directed mutagenesis

2013 ◽  
Vol 27 (S1) ◽  
Author(s):  
Endia Blunt ◽  
Madhusoodanan Mottamal ◽  
Chantelle Collins ◽  
Kelly Johanson
Keyword(s):  
Dna Binding ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Binding Studies ◽  
Dna Binding Studies

scholarly journals Distinct sites for myotoxic and membrane-damaging activities in the C-terminal region of a Lys49-phospholipase A2

2002 ◽  
Vol 366 (3) ◽  
pp. 971-976 ◽  
Author(s):  
Lucimara CHIOATO ◽  
Arthur H.C. de OLIVEIRA ◽  
Roberto RULLER ◽  
Juliana M. SÁ ◽  
Richard J. WARD
Keyword(s):  
Phospholipase A2 ◽  
Lipid Membrane ◽  
Terminal Region ◽  
Site Directed Mutagenesis ◽  
Phospholipid Bilayer ◽  
Directed Mutagenesis ◽  
Aromatic Residues ◽  
Loop Region ◽  
Terminal Loop ◽  
Arginine Residues

Bothropstoxin-I (BthTx-I) is a Lys49-phospholipase A2 from the venom of Bothrops jararacussu which demonstrates both myotoxic and Ca2+-independent membrane-damaging activities. The structural determinants of these activities are poorly defined, therefore site-directed mutagenesis has been used to substitute all cationic and aromatic residues between positions 115 and 129 in the C-terminal loop region of the protein. Substitution of lysine and arginine residues with alanine in the region 117—122 resulted in a significant reduction of myotoxic activity of the recombinant BthTx-I. With the exception of Lys122, these same substitutions did not significantly alter the Ca2+-independent membrane-damaging activity. In contrast, substitution of the positively-charged residues at positions 115, 116 and 122 resulted in reduced Ca2+-independent membrane-damaging activity but, with the exception of Lys122, had no effect on myotoxicity. These results indicate that the two activities are independent and are determined by discrete yet partially overlapping motifs in the C-terminal loop. Results from site-directed mutagenesis of the aromatic residues in the same part of the protein suggest that a region including residues 115—119 interacts superficially with the membrane interface and that the residues around position 125 partially insert into the lipid membrane. These results represent the first detailed mapping of a myotoxic site in a phospholipase A2, and support a model of a Ca2+-independent membrane-damaging mechanism in which the C-terminal region of BthTx-I interacts with and contributes to the perturbation of the phospholipid bilayer.

Site-directed mutagenesis studies of the aromatic residues at the active site of a lipase from Malassezia globosa

Biochimie ◽  
2014 ◽  
Vol 102 ◽  
pp. 29-36 ◽  
Author(s):  
Chongliang Gao ◽  
Dongming Lan ◽  
Lu Liu ◽  
Houjin Zhang ◽  
Bo Yang ◽  
...  
Keyword(s):  
Active Site ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Aromatic Residues ◽  
Malassezia Globosa
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