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A Continuous Fluorimetric Assay for ATPase Activity

Overview
Journal Biochem J
Specialty Biochemistry
Date 1990 Mar 1
PMID 2138458
Citations 11
Authors
U Banik
S Roy
Affiliations
Soon will be listed here.
Abstract

A new continuous coupled fluorimetric assay is described for ATPases in general. Thus phosphate released from ATP hydrolysis is coupled to the nucleoside phosphorylase reaction using 7-methylguanosine as a fluorescent substrate for the nucleoside phosphorylase reaction. The hydrolysis of 7-methylguanosine leads to 7-methylguanine, which has lower quantum yield and hence can be used to monitor ATP hydrolysis continuously. The method has the potential to be extended to GTPase and nucleotidyltransferase assays.

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References
1.
Driska S, Hartshorne D . The contractile proteins of smooth muscle. Properties and components of a Ca2+-sensitive actomyosin from chicken gizzard. Arch Biochem Biophys. 1975; 167(1):203-12. DOI: 10.1016/0003-9861(75)90457-9. View
2.
Brattin W, Waller R . Calcium inhibition of rat liver microsomal calcium-dependent ATPase. J Biol Chem. 1983; 258(11):6724-9. View
3.
Kulikowska E, Bzowska A, Wierzchowski J, SHUGAR D . Properties of two unusual, and fluorescent, substrates of purine-nucleoside phosphorylase: 7-methylguanosine and 7-methylinosine. Biochim Biophys Acta. 1986; 874(3):355-63. DOI: 10.1016/0167-4838(86)90035-x. View
4.
Anderson K, Murphy A . Alterations in the structure of the ribose moiety of ATP reduce its effectiveness as a substrate for the sarcoplasmic reticulum ATPase. J Biol Chem. 1983; 258(23):14276-8. View
5.
Chan K, Delfert D, Junger K . A direct colorimetric assay for Ca2+ -stimulated ATPase activity. Anal Biochem. 1986; 157(2):375-80. DOI: 10.1016/0003-2697(86)90640-8. View