Actin-inhibition and folding of vertebrate deoxyribonuclease I are affected by mutations at residues 67 and 114

Amino acid (aa) residues (Val-67 and Ala-114) have been suggested as being mainly responsible for actin-binding in human and bovine deoxyribonucleases I (DNase I). This study presents evidence of these two aa mutational mechanisms, not only for actin-binding but also for folding of DNase I in mammal...

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Published in:Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 2006, Vol.143 (1), p.70-75
Main Authors: Fujihara, Junko, Hieda, Yoko, Xue, Yuying, Nakagami, Norihito, Imamura, Shinji, Takayama, Koji, Kataoka, Kaori, Takeshita, Haruo
Format: Article
Language:English
Subjects:
Actins - metabolism
African clawed frog
Amino Acid Sequence
Amino Acid Substitution
Animals
Binding Sites
Deoxyribonuclease I - chemistry
Deoxyribonuclease I - genetics
Deoxyribonuclease I - metabolism
G-actin
Human
Molecular Sequence Data
Mutation
Pig
Protein Folding
Rat snake
Rats
Recombinant DNase I
Sequence Homology, Amino Acid
Species Specificity
Substitution mutant
Swine
Viper snake
Viperidae
Xenopus laevis
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Summary:Amino acid (aa) residues (Val-67 and Ala-114) have been suggested as being mainly responsible for actin-binding in human and bovine deoxyribonucleases I (DNase I). This study presents evidence of these two aa mutational mechanisms, not only for actin-binding but also for folding of DNase I in mammals, reptiles and amphibians. Human and viper snake ( Agkistrodon blomhoffii) enzymes are inhibited by actin, whereas porcine, rat snake ( Elaphe quadrivirgata), and African clawed frog ( Xenopus laevis) enzymes are not. To investigate the role of aa at 67, mutants of rat snake (Ile67Val) and viper snake (Val67Ile) enzymes were constructed. After substitution, the rat snake was inhibited by actin, while the viper snake was not. For the role of aa at 114, mutants of viper snake (Phe114Ala), rat snake (Phe114Ala), African clawed frog (Phe114Ala), and porcine (Ser114Ala/Ser114Phe) enzymes were constructed. Strikingly, the substitute mutants for viper snake, rat snake and African clawed frog expressed no protein. The porcine (Ser114Ala) enzyme was inhibited by actin, but not the porcine (Ser114Phe) enzyme. These results suggest that Val-67 may be essential for actin-binding, that Phe-114 may be related to the folding of DNase I in reptiles and amphibians, and that Ala-114 may be indispensable for actin-binding in mammals.
ISSN:1096-4959
1879-1107
DOI:10.1016/j.cbpb.2005.10.004