Asn141 is essential for DNA recognition by EcoRI restriction endonuclease.

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.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Fritz A,Küster W,Alves J

doi

10.1016/s0014-5793(98)01274-5

subject

Has Abstract

pub_date

1998-10-30 00:00:00

pages

66-70

issue

1-2

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(98)01274-5

journal_volume

438

pub_type

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