Noncovalent Complexes of APS Reductase from M. Tuberculosis: Delineating a Mechanistic Model Using ESI-FTICR MS

Overview

Journal J Am Soc Mass Spectrom

Specialty Chemistry

Date 2006 Oct 7

PMID 17023175

Citations 9

Authors

Hong Gao

Julie Leary

Kate S Carroll

Carolyn R Bertozzi

Huiyi Chen

Affiliations

Soon will be listed here.

Abstract

ESI-FTICR MS was utilized to characterize a 4Fe-4S containing protein Mycobacterium tuberculosis APS reductase. This enzyme catalyzes the reduction of APS to sulfite and AMP with reducing equivalents from the protein cofactor, thioredoxin. Under nondenaturing conditions, a distribution of the apoprotein, a 2Fe-2S intermediate, and the 4Fe-4S holoprotein were observed. Accurate mass measurements indicated an oxidation state of +2 for the 4Fe-4S cluster, with no disulfide bond in the holoenzyme. Gas-phase stability of the 4Fe-4S cluster was investigated using both in-source and collision induced dissociation, which provided information regarding the relative gas-phase binding strength of iron towards protein ligands and inorganic sulfides. Noncovalent complexes of the holoprotein with several ligands, including APS, thioredoxin, and AMP, were also investigated. Calculated values of dissociation constants for the complexes indicate that AMP binds with a higher affinity to the enzyme intermediate than to the free enzyme. The implications of the binary and ternary complexes observed by gas-phase noncovalent interactions in the mechanism of APS reduction are discussed.

Citing Articles

Characterization of [2Fe-2S]-Cluster-Bridged Protein Complexes and Reaction Intermediates by use of Native Mass Spectrometric Methods.

Jia M, Sen S, Wachnowsky C, Fidai I, Cowan J, Wysocki V Angew Chem Int Ed Engl. 2020; 59(17):6724-6728.

PMID: 32031732 PMC: 7170024. DOI: 10.1002/anie.201915615.

Functional Site Discovery in a Sulfur Metabolism Enzyme by Using Directed Evolution.

Paritala H, Palde P, Carroll K Chembiochem. 2016; 17(19):1873-1878.

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Design, synthesis and evaluation of Fe-S targeted adenosine 5'-phosphosulfate reductase inhibitors.

Paritala H, Suzuki Y, Carroll K Nucleosides Nucleotides Nucleic Acids. 2015; 34(3):199-220.

PMID: 25710356 PMC: 4341950. DOI: 10.1080/15257770.2014.978012.

New targets and inhibitors of mycobacterial sulfur metabolism.

Paritala H, Carroll K Infect Disord Drug Targets. 2013; 13(2):85-115.

PMID: 23808874 PMC: 4332622. DOI: 10.2174/18715265113139990022.

Geometric and electrostatic study of the [4Fe-4S] cluster of adenosine-5'-phosphosulfate reductase from broken symmetry density functional calculations and extended X-ray absorption fine structure spectroscopy.

Bhave D, Han W, Pazicni S, Penner-Hahn J, Carroll K, Noodleman L Inorg Chem. 2011; 50(14):6610-25.

PMID: 21678934 PMC: 3134165. DOI: 10.1021/ic200446c.

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