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  • Title: Asn141 is essential for DNA recognition by EcoRI restriction endonuclease.
    Author: Fritz A, Küster W, Alves J.
    Journal: FEBS Lett; 1998 Oct 30; 438(1-2):66-70. PubMed ID: 9821960.
    Abstract:
    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.
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