Insights into the Cnx1E catalyzed MPT-AMP hydrolysis.
Adenosine Monophosphate
/ metabolism
Amino Acid Sequence
Amino Acids
/ metabolism
Arabidopsis
/ metabolism
Arabidopsis Proteins
/ metabolism
Catalysis
Catalytic Domain
Coenzymes
/ metabolism
Hydrolysis
Metalloproteins
/ metabolism
Molybdenum
/ metabolism
Molybdenum Cofactors
Organophosphorus Compounds
/ metabolism
Pteridines
/ metabolism
Pterins
/ metabolism
Adenylated molybdopterin
Enzyme catalyzed hydrolysis
Molybdenum cofactor
Molybdenum insertase
Journal
Bioscience reports
ISSN: 1573-4935
Titre abrégé: Biosci Rep
Pays: England
ID NLM: 8102797
Informations de publication
Date de publication:
31 01 2020
31 01 2020
Historique:
received:
29
05
2019
revised:
17
12
2019
accepted:
19
12
2019
pubmed:
21
12
2019
medline:
1
12
2020
entrez:
21
12
2019
Statut:
ppublish
Résumé
Molybdenum insertases (Mo-insertases) catalyze the final step of molybdenum cofactor (Moco) biosynthesis, an evolutionary old and highly conserved multi-step pathway. In the first step of the pathway, GTP serves as substrate for the formation of cyclic pyranopterin monophosphate, which is subsequently converted into molybdopterin (MPT) in the second pathway step. In the following synthesis steps, MPT is adenylated yielding MPT-AMP that is subsequently used as substrate for enzyme catalyzed molybdate insertion. Molybdate insertion and MPT-AMP hydrolysis are catalyzed by the Mo-insertase E-domain. Earlier work reported a highly conserved aspartate residue to be essential for Mo-insertase functionality. In this work, we confirmed the mechanistic relevance of this residue for the Arabidopsis thaliana Mo-insertase Cnx1E. We found that the conservative substitution of Cnx1E residue Asp274 by Glu (D274E) leads to an arrest of MPT-AMP hydrolysis and hence to the accumulation of MPT-AMP. We further showed that the MPT-AMP accumulation goes in hand with the accumulation of molybdate. By crystallization and structure determination of the Cnx1E variant D274E, we identified the potential reason for the missing hydrolysis activity in the disorder of the region spanning amino acids 269 to 274. We reasoned that this is caused by the inability of a glutamate in position 274 to coordinate the octahedral Mg2+-water complex in the Cnx1E active site.
Identifiants
pubmed: 31860061
pii: 221653
doi: 10.1042/BSR20191806
pmc: PMC6954367
pii:
doi:
Substances chimiques
Amino Acids
0
Arabidopsis Proteins
0
Coenzymes
0
Metalloproteins
0
Molybdenum Cofactors
0
Organophosphorus Compounds
0
Pteridines
0
Pterins
0
nulibry
0
molybdate
14259-85-9
Adenosine Monophosphate
415SHH325A
Molybdenum
81AH48963U
molybdenum cofactor
ATN6EG42UQ
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
© 2020 The Author(s).
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