Carbocyclic glycinamide ribonucleotide is a substrate for glycinamide ribonucleotide transformylase

Caperelli, Carol A.; Price, Martyn F.

doi:10.1016/0003-9861(88)90602-9

Cited by 18 publications

(2 citation statements)

References 11 publications

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“…(c) GAR Analogues. In accordance with previous observations (Caperelli & Price, 1988) a /3-GAR analogue in which the Cl'-C4' bridging oxygen was replaced with a methylene group was found to be a substrate for the E. coli enzyme. The Km of this carbocyclic GAR analogue was approximately 2-fold higher than the Km of /3-GAR, with the km being roughly the same.…”

Section: Resultssupporting

confidence: 92%

Subcloning, characterization, and affinity labeling of Escherichia coli glycinamide ribonucleotide transformylase

Inglese¹,

Dl²,

Shiau³

et al. 1990

Biochemistry

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Glycinamide ribonucleotide transformylase (GAR TFase; EC 2.1.2.2) has been purified 70-fold to apparent homogeneity from Escherichia coli harboring an expression vector encoding the purN gene product, GAR TFase. The protein is a monomer of Mr 23,241 and catalyzes a single reaction. Steady-state kinetic parameters for the enzyme have been obtained. The structural requirements for cofactor utilization have been investigated and found to parallel those of the multifunctional avian enzyme. The enzyme was inactivated with the affinity label N10-(bromoacetyl)-5,8-dideazafolate in a stoichiometric and active-site-specific manner. The ionization state of the cofactor analogue in the enzyme-cofactor complex appears to require the dissociation of the proton at N3 of the pyrimidine within the complex.

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Section: Resultssupporting

confidence: 92%

Subcloning, characterization, and affinity labeling of Escherichia coli glycinamide ribonucleotide transformylase

Inglese¹,

Dl²,

Shiau³

et al. 1990

Biochemistry

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“…The glycine moiety of the β-GAR still had weak density, but the well-defined electron density around the anomeric carbon (C1) of GAR gave no indication of additional binding of the α-anomer. This interpretation is in agreement with kinetic studies that show only β-GAR is utilized by GAR Tfase and that the presence of α-GAR in the anomeric mixture has no effect on enzyme activity ( , ). InsightII (MSI, San Diego, CA) was used to generate and minimize coordinates for 10-formyl-TDAF, and the topology and parameter files required by X-PLOR were generated by XPLO2D ().…”

Section: Methodssupporting

confidence: 91%

New Insights into Inhibitor Design from the Crystal Structure and NMR Studies ofEscherichia coliGAR Transformylase in Complex with β-GAR and 10-Formyl-5,8,10-trideazafolic Acid^,

et al. 1999

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The crystal structure of Escherichia coli GAR Tfase at 2.1 A resolution in complex with 10-formyl-5,8,10-trideazafolic acid (10-formyl-TDAF, K(i) = 260 nM), an inhibitor designed to form an enzyme-assembled multisubstrate adduct with the substrate, beta-GAR, was studied to determine the exact nature of its inhibitory properties. Rather than forming the expected covalent adduct, the folate inhibitor binds as the hydrated aldehyde (gem-diol) in the enzyme active site, in a manner that mimics the tetrahedral intermediate of the formyl transfer reaction. In this hydrated form, the inhibitor not only provides unexpected insights into the catalytic mechanism but also explains the 10-fold difference in inhibitor potency between 10-formyl-TDAF and the corresponding alcohol, and a further 10-fold difference for inhibitors that lack the alcohol. The presence of the hydrated aldehyde was confirmed in solution by (13)C-(1)H NMR spectroscopy of the ternary GAR Tfase-beta-GAR-10-formyl-TDAF complex using the (13)C-labeled 10-formyl-TDAF. This insight into the behavior of the inhibitor, which is analogous to protease or transaminase inhibitors, provides a novel and previously unrecognized basis for the design of more potent inhibitors of the folate-dependent formyl transfer enzymes of the purine biosynthetic pathway and development of anti-neoplastic agents.

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(±)‐5′‐Nor ribofuranoside carbocyclic guanosine

Patil

Schneller

1991

Journal of Heterocyclic Chem

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Carbocyclic glycinamide ribonucleotide is a substrate for glycinamide ribonucleotide transformylase

Cited by 18 publications

References 11 publications

Subcloning, characterization, and affinity labeling of Escherichia coli glycinamide ribonucleotide transformylase

Subcloning, characterization, and affinity labeling of Escherichia coli glycinamide ribonucleotide transformylase

New Insights into Inhibitor Design from the Crystal Structure and NMR Studies ofEscherichia coliGAR Transformylase in Complex with β-GAR and 10-Formyl-5,8,10-trideazafolic Acid^,

(±)‐5′‐Nor ribofuranoside carbocyclic guanosine

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Carbocyclic glycinamide ribonucleotide is a substrate for glycinamide ribonucleotide transformylase

Cited by 18 publications

References 11 publications

Subcloning, characterization, and affinity labeling of Escherichia coli glycinamide ribonucleotide transformylase

Subcloning, characterization, and affinity labeling of Escherichia coli glycinamide ribonucleotide transformylase

New Insights into Inhibitor Design from the Crystal Structure and NMR Studies ofEscherichia coliGAR Transformylase in Complex with β-GAR and 10-Formyl-5,8,10-trideazafolic Acid,

(±)‐5′‐Nor ribofuranoside carbocyclic guanosine

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Resources

About

New Insights into Inhibitor Design from the Crystal Structure and NMR Studies ofEscherichia coliGAR Transformylase in Complex with β-GAR and 10-Formyl-5,8,10-trideazafolic Acid^,