TY - JOUR
T1 - Electrolysis-mediated irreversible inactivation of lipoxygenase directed toward electroaffinity labelling
AU - Holmes, Thomas J.
AU - Vennerstrom, Jonathan L.
AU - John, Varghese
N1 - Funding Information:
The generous financial support of the U.S. National Institutes of Health via New Investigator Award(T.J.H., Jr.) No. R23-29844 is gratefully acknowledged. The technical assistance of Professor Larry Miller, Department of Chemistry, University Minnesota and Professor Ron Blankespoor, Calvin College, Grand Rapids, is greatly appreciated as well.
PY - 1984/8/30
Y1 - 1984/8/30
N2 - Irreversible inhibition of soybean lipoxygenase-1 (SL-1) was accomplished via a controlled potential oxidative electrolysis of 1,5-dihydroxynaphthalene (1,5-DHN) at +0.8 V vs SCE. The inactivation of SL-1 with this known inhibitor was greatly enhanced under these electrolytic conditions to which the enzyme itself was stable. Electrolyses were run at 0°C in a 0.05 M phosphate buffer, pH 7.0, using graphite cloth electrodes. The rate of inactivation was observed to be limited by and dependent on the anodic oxidation of 1,5-DHN. The non-oxidizable (at this potential) inhibitor indomethacin was shown to protect the enzyme from irreversible inactivation, however, an external nucleophile (2-mercaptoethanol) had little effect. These initial studies support the capability of such electrochemical methods for the site-specific covalent modification (affinity labelling) of lipoxygenase, and perhaps other enzymes.
AB - Irreversible inhibition of soybean lipoxygenase-1 (SL-1) was accomplished via a controlled potential oxidative electrolysis of 1,5-dihydroxynaphthalene (1,5-DHN) at +0.8 V vs SCE. The inactivation of SL-1 with this known inhibitor was greatly enhanced under these electrolytic conditions to which the enzyme itself was stable. Electrolyses were run at 0°C in a 0.05 M phosphate buffer, pH 7.0, using graphite cloth electrodes. The rate of inactivation was observed to be limited by and dependent on the anodic oxidation of 1,5-DHN. The non-oxidizable (at this potential) inhibitor indomethacin was shown to protect the enzyme from irreversible inactivation, however, an external nucleophile (2-mercaptoethanol) had little effect. These initial studies support the capability of such electrochemical methods for the site-specific covalent modification (affinity labelling) of lipoxygenase, and perhaps other enzymes.
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U2 - 10.1016/0006-291X(84)90393-0
DO - 10.1016/0006-291X(84)90393-0
M3 - Article
C2 - 6433914
AN - SCOPUS:0021129997
VL - 123
SP - 156
EP - 162
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -