TY - JOUR
T1 - Evidence that oxidative dephosphorylation by the nonheme Fe(II), α-ketoglutarate:UMP oxygenase occurs by stereospecific hydroxylation
AU - Goswami, Anwesha
AU - Liu, Xiaodong
AU - Cai, Wenlong
AU - Wyche, Thomas P.
AU - Bugni, Tim S.
AU - Meurillon, Maïa
AU - Peyrottes, Suzanne
AU - Perigaud, Christian
AU - Nonaka, Koichi
AU - Rohr, Jürgen
AU - Van Lanen, Steven G.
N1 - Publisher Copyright:
© 2017 Federation of European Biochemical Societies
PY - 2017/2/1
Y1 - 2017/2/1
N2 - LipL and Cpr19 are nonheme, mononuclear Fe(II)-dependent, α-ketoglutarate (αKG):UMP oxygenases that catalyze the formation of CO2, succinate, phosphate, and uridine-5′-aldehyde, the last of which is a biosynthetic precursor for several nucleoside antibiotics that inhibit bacterial translocase I (MraY). To better understand the chemistry underlying this unusual oxidative dephosphorylation and establish a mechanistic framework for LipL and Cpr19, we report herein the synthesis of two biochemical probes—[1′,3′,4′,5′,5′-2H]UMP and the phosphonate derivative of UMP—and their activity with both enzymes. The results are consistent with a reaction coordinate that proceeds through the loss of one 2H atom of [1′,3′,4′,5′,5′-2H]UMP and stereospecific hydroxylation geminal to the phosphoester to form a cryptic intermediate, (5′R)-5′-hydroxy-UMP. Thus, these enzyme catalysts can additionally be assigned as UMP hydroxylase-phospholyases.
AB - LipL and Cpr19 are nonheme, mononuclear Fe(II)-dependent, α-ketoglutarate (αKG):UMP oxygenases that catalyze the formation of CO2, succinate, phosphate, and uridine-5′-aldehyde, the last of which is a biosynthetic precursor for several nucleoside antibiotics that inhibit bacterial translocase I (MraY). To better understand the chemistry underlying this unusual oxidative dephosphorylation and establish a mechanistic framework for LipL and Cpr19, we report herein the synthesis of two biochemical probes—[1′,3′,4′,5′,5′-2H]UMP and the phosphonate derivative of UMP—and their activity with both enzymes. The results are consistent with a reaction coordinate that proceeds through the loss of one 2H atom of [1′,3′,4′,5′,5′-2H]UMP and stereospecific hydroxylation geminal to the phosphoester to form a cryptic intermediate, (5′R)-5′-hydroxy-UMP. Thus, these enzyme catalysts can additionally be assigned as UMP hydroxylase-phospholyases.
KW - antibiotic
KW - biosynthesis
KW - nonheme iron
KW - nucleoside
KW - oxygenase
KW - translocase I
UR - http://www.scopus.com/inward/record.url?scp=85012982886&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85012982886&partnerID=8YFLogxK
U2 - 10.1002/1873-3468.12554
DO - 10.1002/1873-3468.12554
M3 - Article
C2 - 28074470
AN - SCOPUS:85012982886
SN - 0014-5793
VL - 591
SP - 468
EP - 478
JO - FEBS Letters
JF - FEBS Letters
IS - 3
ER -