PMID: 6160464Jun 11, 1980Paper

Binding of non-substrate nucleotides to a restriction endonuclease: a model for the interaction of bam HI with its recognition sequence

Nucleic Acids Research
B HinschM R Kula

Abstract

The kinetic constants of the site-specific endonuclease Bam HI for various substrates were determined and binding of non-substrate nucleotides to the enzyme was studied. Agarose gel assays in combination with an integrated Michaelis-Menten equation were used for the evaluation of data. The turnover number was 2.2 min-1 at 37 degrees C with pJC80 DNA as the substrate. It depends on the conformation and base composition of the substrate. Michaelis constants also depend on substrate conformation. Non-substrate polynucleotides were found to inhibit Bam competitively with KI ranging from 10(-6) to > 10(-3) M depending on base composition, base pairing, and helix conformation. Dinucleotides showed sequence-specific, competitive inhibition with KIs ranging from 10(-5) to > 10(-3) M. Mononucleotides and -nucleosides acted noncompetitively. Binding was influenced by the extent of phosphorylation, but not by the nature of the base. KIs varied between 10(-3) and 10(-2) M. The results are discussed with respect to the recognition requirements of Bam HI.

References

Jan 1, 1976·Annual Review of Biochemistry·T M Jovin
Mar 1, 1977·Analytical Biochemistry·A PrunellB Leblanc
Dec 1, 1977·Nucleic Acids Research·M B Mann, H O Smith
Oct 5, 1978·Journal of Molecular Biology·S HattmanA Gottehrer
Feb 2, 1976·Molecular & General Genetics : MGG·H R SmithE S Anderson
Dec 13, 1977·Biochemistry·C Nakai, W Glinsmann
Jul 1, 1976·Nucleic Acids Research·V Pirrotta
Mar 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·N C SeemanA Rich
Jan 25, 1975·Journal of Molecular Biology·M Thomas, R W Davis
Jan 25, 1980·Nucleic Acids Research·J L Woodhead, A D Malcolm
Apr 1, 1964·Journal of Molecular Biology·R B INMAN, R L BALDWIN

❮ Previous
Next ❯

Citations

Jul 10, 1981·Nucleic Acids Research·B Hinsch, M R Kula
Nov 30, 2000·Proceedings of the National Academy of Sciences of the United States of America·J B BigginsJ S Thorson

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bacteriophage: Phage Therapy

Phage therapy uses bacterial viruses (bacteriophages) to treat bacterial infections and is widely being recognized as an alternative to antibiotics. Here is the latest research.