Pronucleotide Probes Reveal a Diverging Specificity for AMPylation vs UMPylation of Human and Bacterial Nucleotide Transferases
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Pronucleotide Probes Reveal a Diverging Specificity for AMPylation vs UMPylation of Human and Bacterial Nucleotide Transferases. / Mostert, Dietrich; Bubeneck, Wilhelm Andrei; Rauh, Theresa; Kielkowski, Pavel; Itzen, Aymelt; Jung, Kirsten; Sieber, Stephan A.
in: BIOCHEMISTRY-US, Jahrgang 63, Nr. 5, 05.03.2024, S. 651-659.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Pronucleotide Probes Reveal a Diverging Specificity for AMPylation vs UMPylation of Human and Bacterial Nucleotide Transferases
AU - Mostert, Dietrich
AU - Bubeneck, Wilhelm Andrei
AU - Rauh, Theresa
AU - Kielkowski, Pavel
AU - Itzen, Aymelt
AU - Jung, Kirsten
AU - Sieber, Stephan A
PY - 2024/3/5
Y1 - 2024/3/5
N2 - AMPylation is a post-translational modification utilized by human and bacterial cells to modulate the activity and function of specific proteins. Major AMPylators such as human FICD and bacterial VopS have been studied extensively for their substrate and target scope in vitro. Recently, an AMP pronucleotide probe also facilitated the in situ analysis of AMPylation in living cells. Based on this technology, we here introduce a novel UMP pronucleotide probe and utilize it to profile uninfected and Vibrio parahaemolyticus infected human cells. Mass spectrometric analysis of labeled protein targets reveals an unexpected promiscuity of human nucleotide transferases with an almost identical target set of AMP- and UMPylated proteins. Vice versa, studies in cells infected by V. parahaemolyticus and its effector VopS revealed solely AMPylation of host enzymes, highlighting a so far unknown specificity of this transferase for ATP. Taken together, pronucleotide probes provide an unprecedented insight into the in situ activity profile of crucial nucleotide transferases, which can largely differ from their in vitro activity.
AB - AMPylation is a post-translational modification utilized by human and bacterial cells to modulate the activity and function of specific proteins. Major AMPylators such as human FICD and bacterial VopS have been studied extensively for their substrate and target scope in vitro. Recently, an AMP pronucleotide probe also facilitated the in situ analysis of AMPylation in living cells. Based on this technology, we here introduce a novel UMP pronucleotide probe and utilize it to profile uninfected and Vibrio parahaemolyticus infected human cells. Mass spectrometric analysis of labeled protein targets reveals an unexpected promiscuity of human nucleotide transferases with an almost identical target set of AMP- and UMPylated proteins. Vice versa, studies in cells infected by V. parahaemolyticus and its effector VopS revealed solely AMPylation of host enzymes, highlighting a so far unknown specificity of this transferase for ATP. Taken together, pronucleotide probes provide an unprecedented insight into the in situ activity profile of crucial nucleotide transferases, which can largely differ from their in vitro activity.
KW - Humans
KW - Nucleotides/metabolism
KW - Transferases/metabolism
KW - Bacterial Proteins/chemistry
KW - Adenosine Monophosphate/metabolism
KW - Protein Processing, Post-Translational
U2 - 10.1021/acs.biochem.3c00568
DO - 10.1021/acs.biochem.3c00568
M3 - SCORING: Journal article
C2 - 38388156
VL - 63
SP - 651
EP - 659
JO - BIOCHEMISTRY-US
JF - BIOCHEMISTRY-US
SN - 0006-2960
IS - 5
ER -
