FEBS Letters | |
Asn141 is essential for DNA recognition by EcoRI restriction endonuclease | |
Fritz, Andreas2  Küster, Wolfgang1  Alves, Jürgen1  | |
[1] Zentrum Biochemie, Medizinische Hochschule Hannover, Carl-Neuberg-Str. 1, D-30625 Hannover, Germany;Institut für Pathologie, GSG-Forschungszentrum für Umwelt und Gesundheit GmbH, Ingolstädter Landstr. 1, D-85758 Oberschleißheim, Germany | |
关键词: DNA-protein interaction; Type II restriction enzyme; Sequence specificity; Site-directed mutagenesis; Restriction modification; Hydrogen bond; CD; circular dichroism; | |
DOI : 10.1016/S0014-5793(98)01274-5 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
The amino acid residue Asn141 of the restriction endonuclease EcoRI was proposed to make three hydrogen bonds to both adenine residues within the recognition sequence -GAATTC-. We have mutated Asn141 to alanine, aspartate, serine, and tyrosine. Only the serine mutant is active under normal buffer conditions although 1000-fold less than wild-type EcoRI. The alanine and aspartate mutants can be activated by Mn2+. At acidic pH the latter mutant becomes even more active than the wild-type enzyme in the presence of Mn2+. We conclude that Asn141 is essential for DNA recognition and that serine can partly substitute it.
【 授权许可】
Unknown
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