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Reversible regulation of the nitrogenase iron protein from Rhodospirillum rubrum by ADP-ribosylation in vitro.
TLDR
NAD-dependent modification and concomitant inactivation of the Fe protein were demonstrated in crude extracts of R. rubrum and it was shown that the inactivating enzyme activity is stimulated by divalent metal ions and ADP, that O2-denatured Fe protein will not serve as a substrate, and that dithionite inhibits the modification reaction. Expand
Two Gα i1 Rate-Modifying Mutations Act in Concert to Allow Receptor-Independent, Steady-State Measurements of RGS Protein Activity
TLDR
To enable nonradioactive, homogeneous detection of RGS protein effects on Gαi1(R178M/A326S), the authors developed a Transcreener ® fluorescence polarization immunoassay based on a monoclonal antibody that recognizes GDP with greater than 100-fold selectivity over GTP. Expand
Purification and properties of dinitrogenase reductase ADP-ribosyltransferase from the photosynthetic bacterium Rhodospirillum rubrum.
TLDR
The enzyme that catalyzes the ADP-ribosylation and concomitant inactivation of dinitrogenase reductase in Rhodospirillum rubrum has been purified greater than 19,000-fold to near homogeneity and DRAT is proposed as the working name for the enzyme. Expand
Genes coding for the reversible ADP-ribosylation system of dinitrogenase reductase from Rhodospirillum rubrum
TLDR
The cloning and characterization of the genes encoding the ADP-ribosyltransferase (draT) and theADP- ribosylglycohydrolase ( draG) involved in this regulation of nitrogen fixation activity in Rhodospirillum rubrum are described. Expand
A High-Throughput Assay for Rho Guanine Nucleotide Exchange Factors Based on the Transcreener GDP Assay
TLDR
A simple, generic biochemical assay method for measuring GEF activity based on the fact that GDP dissociation is generally the rate-limiting step in the Rho GTPase catalytic cycle, and thus addition of a GEF causes an increase in steady-state GTP enzyme activity is developed. Expand
Evaluating Modulators of “Regulator of G‐protein Signaling” (RGS) Proteins
TLDR
High‐throughput screening procedures for identifying modulators of RGS protein‐mediated GTPase acceleration (GAP activity), for assessment of R GS domain/Gα interactions, and for validation of candidate GAP‐modulatory molecules with the single‐turnover GTP hydrolysis assay are described. Expand
Substitution of histidine for arginine-101 of dinitrogenase reductase disrupts electron transfer to dinitrogenase.
TLDR
The interaction of dinitrogenase reductase with dinitrogensase during reductant-independent ATP hydrolysis is different than the interaction between the two proteins during electron transfer; the substitution of histidine for arginine at position 101 disrupts only the latter interaction. Expand
ADP-ribosylation of dinitrogenase reductase from Clostridium pasteurianum prevents its inhibition of nitrogenase from Azotobacter vinelandii.
TLDR
ADP-ribosylation inactivated Cp2 and prevented its formation of a tight complex with dinitrogenase from Azotobacter vinelandii (Av1) and the complex between Cp1 and Av1 could not be ADP- ribosylated once it formed. Expand
Effect of nucleotides on the activity of dinitrogenase reductase ADP-ribosyltransferase from Rhodospirillum rubrum.
TLDR
It is concluded that MgADP stimulates DRAT activity by lowering the Km for NAD and that M gADP exerts its effect by binding to dinitrogenase reductase. Expand
Characterization and optimization of a red-shifted fluorescence polarization ADP detection assay.
TLDR
Development of antibodies with >100-fold selectivity for ADP versus ATP are described, which enable robust detection of initial velocity rates at ATP concentrations ranging from 0.1 microM to 1,000 microM in a competitive fluorescence polarization (FP) immunoassay. Expand
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