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Folding topology and DNA binding of the N‐terminal fragment of ada protein
Author(s) -
Sakashita Hitoshi,
Sakuma Takahiko,
Ohkubo Tadayasu,
Kainosho Masatsune,
Sakumi Kunihiko,
Sekiguchi Mutsuo,
Morikawa Kosuke
Publication year - 1993
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 - 10.1016/0014-5793(93)81351-y
Subject(s) - plasmid , escherichia coli , fragment (logic) , dna , microbiology and biotechnology , chemistry , molecule , duplex (building) , biology , biochemistry , stereochemistry , gene , organic chemistry , computer science , programming language
Three amino terminal fragments of Escherichia coli Ada protein (39 kDa) with different molecular masses (14 kDa, 16 kDa and 20 kDa) were prepared in large quantities from an E. coli strain harboring plasmids constructed for the overproduction of the truncated proteins. The three fragments can be methylated to an extent similar to that of the intact molecule. The methylated 16 kDa fragment specifically binds to the ada box on a DNA duplex. NMR analyses revealed that the 14 kDa fragment comprises two α‐helices and a β‐sheet with parallel and anti‐parallel mixed strands. A comparison of the 15 N‐ 1 H HMQC spectra of the fragments has led to the conclusion that this tertiary structure within the 14 kDa fragment is retained in the larger 16 kDa and 20 kDa fragments.