Premium
Genetic engineering, production and characterisation of monomeric variants of the dimeric Serratia marcescens endonuclease
Author(s) -
Franke Ingo,
Meiss Gregor,
Blecher Dinah,
Gimadutdinow Oleg,
Urbanke Claus,
Pingoud Alfred
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00279-8
Subject(s) - nuclease , endonuclease , micrococcal nuclease , dna , dimer , monomer , serratia marcescens , chemistry , enzyme , biochemistry , stereochemistry , biology , escherichia coli , gene , organic chemistry , nucleosome , histone , polymer
The Serratia nuclease is a non‐specific endonuclease which cleaves single‐ and double‐stranded RNA and DNA. It is a member of a large family of related endonucleases, most of which are dimers of identical subunits, with the notable exception of the Anabaena nuclease which is a monomer. In order to find out whether the dimer state of the Serratia nuclease is essential for its function we have produced variants of this nuclease which based on the crystal structure (Miller, M.D. and Krause, K.L. (1996), Protein Science 5, 24–33) were expected to be unable to dimerise. We demonstrate here that these variants, H184A, H184N, H184T and H184R, are monomers and have the same secondary structure, stability towards chemical denaturation and activity as the wild‐type enzyme. This allows to conclude that the dimeric state is not essential for the catalytic function of the Serratia nuclease. In contrast, the S179C variant which is also a monomer shows little activity, presumably because this amino acid substitution changes the structure of the enzyme.