TY - JOUR
T1 - The Arabidopsis thalianaK+-uptake permease 7 (AtKUP7) contains a functional cytosolic adenylate cyclase catalytic centre
AU - Al-Younis, Inas
AU - Wong, Aloysius Tze
AU - Gehring, Christoph A
N1 - KAUST Repository Item: Exported on 2020-10-01
Acknowledgements: This project was funded by King Abdullah University of Science and Technology.
PY - 2015/11/27
Y1 - 2015/11/27
N2 - Adenylate Cyclases (ACs) catalyze the formation of the second messenger cyclic adenosine 3′, 5′-monophosphate (cAMP) from adenosine 5’-triphosphate (ATP). Although cAMP is increasingly recognized as an important signaling molecule in higher plants, ACs have remained somewhat elusive. Here we used a search motif derived from experimentally tested guanylyl cyclases (GCs), substituted the residues essential for substrate specificity and identified the Arabidopsis thaliana K+-uptake permease 7 (AtKUP7) as one of several candidate ACs. Firstly, we show that a recombinant N-terminal, cytosolic domain of AtKUP71-100 is able to complement the AC-deficient mutant cyaA in Escherichia coli and thus restoring the fermentation of lactose, and secondly, we demonstrate with both enzyme immunoassays and mass spectrometry that a recombinant AtKUP71-100 generates cAMP in vitro.
AB - Adenylate Cyclases (ACs) catalyze the formation of the second messenger cyclic adenosine 3′, 5′-monophosphate (cAMP) from adenosine 5’-triphosphate (ATP). Although cAMP is increasingly recognized as an important signaling molecule in higher plants, ACs have remained somewhat elusive. Here we used a search motif derived from experimentally tested guanylyl cyclases (GCs), substituted the residues essential for substrate specificity and identified the Arabidopsis thaliana K+-uptake permease 7 (AtKUP7) as one of several candidate ACs. Firstly, we show that a recombinant N-terminal, cytosolic domain of AtKUP71-100 is able to complement the AC-deficient mutant cyaA in Escherichia coli and thus restoring the fermentation of lactose, and secondly, we demonstrate with both enzyme immunoassays and mass spectrometry that a recombinant AtKUP71-100 generates cAMP in vitro.
UR - http://hdl.handle.net/10754/621444
UR - http://doi.wiley.com/10.1016/j.febslet.2015.11.038
UR - http://www.scopus.com/inward/record.url?scp=84959555428&partnerID=8YFLogxK
U2 - 10.1016/j.febslet.2015.11.038
DO - 10.1016/j.febslet.2015.11.038
M3 - Article
C2 - 26638082
SN - 0014-5793
VL - 589
SP - 3848
EP - 3852
JO - FEBS Letters
JF - FEBS Letters
IS - 24PartB
ER -