New insight into the transcarbamylase family: the structure of putrescine transcarbamylase, a key catalyst for fermentative utilization of agmatine.

PloS One
Luis Mariano PoloVicente Rubio

Abstract

Transcarbamylases reversibly transfer a carbamyl group from carbamylphosphate (CP) to an amine. Although aspartate transcarbamylase and ornithine transcarbamylase (OTC) are well characterized, little was known about putrescine transcarbamylase (PTC), the enzyme that generates CP for ATP production in the fermentative catabolism of agmatine. We demonstrate that PTC (from Enterococcus faecalis), in addition to using putrescine, can utilize L-ornithine as a poor substrate. Crystal structures at 2.5 Å and 2.0 Å resolutions of PTC bound to its respective bisubstrate analog inhibitors for putrescine and ornithine use, N-(phosphonoacetyl)-putrescine and δ-N-(phosphonoacetyl)-L-ornithine, shed light on PTC preference for putrescine. Except for a highly prominent C-terminal helix that projects away and embraces an adjacent subunit, PTC closely resembles OTCs, suggesting recent divergence of the two enzymes. Since differences between the respective 230 and SMG loops of PTC and OTC appeared to account for the differential preference of these enzymes for putrescine and ornithine, we engineered the 230-loop of PTC to make it to resemble the SMG loop of OTCs, increasing the activity with ornithine and greatly decreasing the activity with put...Continue Reading

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Citations

Aug 15, 2015·International Journal of Molecular Sciences·Dashuang ShiMendel Tuchman
Apr 20, 2020·The FEBS Journal·Tjeerd PolsBert Poolman

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Datasets Mentioned

BETA
E8M734

Methods Mentioned

BETA
X-ray
PISA
size exclusion chromatography

Software Mentioned

MOLREP CCP4 suite
Pymol
MOLREP
PISA
TRUNCATE CCP4
PROCHECK
hOTC
GraphPad Prism
SSM
REFMAC5

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