Hostname: page-component-848d4c4894-8bljj Total loading time: 0 Render date: 2024-07-02T17:27:04.803Z Has data issue: false hasContentIssue false
  • English
  • Français

The novel fluorescent CDP-analogue (Pβ)MABA-CDP is a specific probe for the NMP binding site of UMP/CMP kinase

Published online by Cambridge University Press:  25 April 2001

MARKUS G. RUDOLPH ,
THOMAS J.H. VEIT  and
JOCHEN REINSTEIN
MARKUS G. RUDOLPH
Affiliation:
Max-Planck-Institut für Molekulare Physiologie, Abetilung Physikalische Biochemie, Otto-Hahn-Strasse 11, 44227 Dormund, Germany
THOMAS J.H. VEIT
Affiliation:
Max-Planck-Institut für Molekulare Physiologie, Abetilung Physikalische Biochemie, Otto-Hahn-Strasse 11, 44227 Dormund, Germany
JOCHEN REINSTEIN
Affiliation:
Max-Planck-Institut für Molekulare Physiologie, Abetilung Physikalische Biochemie, Otto-Hahn-Strasse 11, 44227 Dormund, Germany
Get access

Abstract

Direct thermodynamic and kinetic investigations of the binding of nucleotides to the nucleoside monophosphate (NMP) site of NMP kinases have not been possible so far because a spectroscopic probe was not available. By coupling a fluorescent N-methylanthraniloyl- (mant) group to the β-phosphate of CDP via a butyl linker, a CDP analogue [(Pβ)MABA-CDP] was obtained that still binds specifically to the NMP site of UmpKdicty, because the base and the ribose moieties, which are involved in specific interactions, are not modified. This allows the direct determination of binding constants for its substrates in competition experiments.

Keywords

fluorescencekinaseskineticsnucleotide analogues
Type
Research Article
Information
Protein Science , Volume 8 , Issue 12 , December 1999 , pp. 2697 - 2704
Copyright
© 1999 The Protein Society

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)