Purification and characterization of the restriction endonuclease RsrI, an isoschizomer of EcoRI.
Rhodobacter sphaeroides strain 630 produces restriction enzyme RsrI which is an isoschizomer of EcoRI. We have purified this enzyme and initiated a comparison with the EcoRI endonuclease. The properties of RsrI are consistent with a reaction mechanism similar to that of EcoRI: the position of cleavage within the -GAATTC-site is identical, the MgCl2 optimum for the cleavage is identical, and the pH profile is similar. Methylation of the substrate sequence by the EcoRI methylase protects the site from cleavage by the RsrI endonuclease. RsrI cross-reacts strongly with anti-EcoRI serum indicating three-dimensional structural similarities. We have determined the sequence of 34 N terminal amino acids for RsrI and this sequence possesses significant similarity to the EcoRI N terminus.
Greene
P J
PJ
Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0554.
Ballard
B T
BT
Stephenson
F
F
Kohr
W J
WJ
Rodriguez
H
H
Rosenberg
J M
JM
Boyer
H W
HW
D000595:Amino Acid Sequence; D002845:Chromatography; D002852:Chromatography, Ion Exchange; D015246:Deoxyribonuclease EcoRI / Q000302:isolation & purification*; D017886:Durapatite; D006882:Hydroxyapatites; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D012242:Rhodobacter sphaeroides / Q000201:enzymology*
D006882:Hydroxyapatites; D007527:Isoenzymes; D017886:Durapatite; D015246:Deoxyribonuclease EcoRI