title: Different Enzymatic Processing of γ-Phosphoramidate and γ-Phosphoester-Modified ATP Analogues.
abstract: Monitoring the activity of ATP-consuming enzymes provides the basis for elucidating their modes of action and regulation. Although a number of ATP analogues have been developed for this, their scope is restricted because of the limited acceptance by respective enzymes. In order to clarify which kind of phosphate-modified ATP analogues are accepted by the α-β-phosphoanhydride-cleaving ubiquitin-activating enzyme 1 (UBA1) and the β-γ-phosphoanhydride-cleaving focal adhesion kinase (FAK), we tested phosphoramidate- and phosphoester-modified ATP analogues. UBA1 and FAK were able to convert phosphoramidate-modified ATP analogues, even with a bulky modification like biotin. In contrast, a phosphoester-modified analogue was poorly accepted. These results demonstrate that minor variations in the design of ATP analogues for monitoring ATP utilization have a significant impact on enzymatic acceptance.
authors.affiliation: Department of Chemistry, Konstanz Research School Chemical Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.
authors.forename: Susanne
authors.initials: S
authors.lastname: Ermert
authors.affiliation: Department of Chemistry, Konstanz Research School Chemical Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.
authors.forename: Stephan M
authors.initials: SM
authors.lastname: Hacker
authors.affiliation: Department of Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Buntru
authors.affiliation: Department of Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Scheffner
authors.affiliation: Department of Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.
authors.forename: Christof R
authors.initials: CR
authors.lastname: Hauck
authors.affiliation: Department of Chemistry, Konstanz Research School Chemical Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Marx
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry* / Q000378:metabolism*; D000577:Amides / Q000737:chemistry*; D004798:Enzymes / Q000378:metabolism*; D004952:Esters / Q000737:chemistry*; D015394:Molecular Structure; D010756:Phosphoric Acids / Q000737:chemistry*; D013379:Substrate Specificity; D054875:Ubiquitination
chemical_list: D000577:Amides; D004798:Enzymes; D004952:Esters; D010756:Phosphoric Acids; D000255:Adenosine Triphosphate; C011067:phosphoramidic acid
keywords: ATP; ATP analogues; enzymes; kinases; phosphorylation