Structures of benzylsuccinate synthase elucidate roles of accessory subunits in glycyl radical enzyme activation and activity

Funk, Michael A.; Judd, Evan T.; Marsh, E. Neil G.; Elliott, Sean J.; Drennan, Catherine L.

doi:10.1073/pnas.1405983111

Cited by 57 publications

(100 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This is not surprising considering that maintaining the glycyl radical during crystal growth is simply not feasible. Despite this technical detail, the crystal structures for GREs have provided significant insight into enzyme mechanism (3,(22)(23)(24)(25)(26)(27)(28). Numerous computational predictions have also been made based on this structural information (29 -31).…”

Section: Resultsmentioning

confidence: 99%

“…In addition to the CbGD, the overall fold of the RiDD is also more similar to the large subunits of benzyl succinate synthase and 4-hydroxyphenyl acetate decarboxylase (24,40). However, both benzyl succinate synthase and 4-hydroxyphenyl acetate decarboxylase have additional peptide subunits, presenting an additional challenge when moving the glycyl radical loop out of the C-terminal domain and into the active site of their respective activating enzymes (25). Funk et al (25) noticed that contacts between the C-terminal glycyl radical domain and "loop 2" of the N-terminal domain in the CbGD resulted in burying an additional 100 Å.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

1,2-Propanediol Dehydration in Roseburia inulinivorans

LaMattina

Keul

Reitzer

et al. 2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

Glycyl radical enzymes (GREs) represent a diverse superfamily of enzymes that utilize a radical mechanism to catalyze difficult, but often essential, chemical reactions. In this work we present the first biochemical and structural data for a GRE-type diol dehydratase from the organism Roseburia inulinivorans (RiDD). Despite high sequence (48% identity) and structural similarity to the GRE-type glycerol dehydratase from Clostridium butyricum, we demonstrate that the RiDD is in fact a diol dehydratase. In addition, the RiDD will utilize both (S)-1,2-propanediol and (R)-1,2-propanediol as a substrate, with an observed preference for the S enantiomer. Based on the new structural information we developed and successfully tested a hypothesis that explains the functional differences we observe.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

1,2-Propanediol Dehydration in Roseburia inulinivorans

LaMattina

Keul

Reitzer

et al. 2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…The structure of fumarate-bound BSS␣␥ was solved initially at 2.3 Å resolution by molecular replacement using the Phenix implementation of Phaser (27) with the previously solved structure of BSS␣␥ as a model (23). Manual rebuilding of the model was performed in Coot (28) followed by refinement with phenix.refine (29) at 2.0 Å resolution.…”

Section: Methodsmentioning

confidence: 99%

“…The structure of toluene-and fumarate-bound BSS␣␤␥ was solved by molecular replacement at 3.30 Å resolution in the Phenix implementation of Phaser using the published 2.00 Å resolution, chloride-bound BSS␣␤␥ structure (23) as an initial model. Due to the low resolution of this dataset, refinement was tightly restrained with the high resolution model as a reference structure in phenix.refine.…”

Section: Methodsmentioning

confidence: 99%

“…Despite the fact that the roles of BSS␤ and BSS␥ are unclear, both proteins are required for viable growth of T. aromatica strain T1 on toluene (21,22). We previously solved structures of the BSS␣␤␥ and BSS␣␥ complexes, revealing that removal of BSS␤ from the complex with BSS␣␥ permits a large conformational change within BSS␣ (23). The core barrel opens, exposing the active site, and a C-terminal domain that harbors the Gly loop (the glycyl radical domain) shifts a few ångströms out of the active site but does not completely vacate the barrel.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Substrate-bound Structures of Benzylsuccinate Synthase Reveal How Toluene Is Activated in Anaerobic Hydrocarbon Degradation

Funk

Marsh

Drennan

2015

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Background: Benzylsuccinate synthase (BSS) catalyzes the formation of a COC bond between toluene and fumarate by a radical mechanism. Results: BSS binds substrates in a buried active site and uses conformational changes to gate access. Conclusion: BSS and related enzymes precisely position substrates for chemistry. Significance: These structures reveal the molecular basis for BSS radical chemistry and substrate specificity.

show abstract

Radical Enzymes

Högbom

Sjöberg

Berggren

2020

Encyclopedia of Life Sciences

View full text Add to dashboard Cite

Radical enzymes harbour a free radical in their polypeptide chain, which participates in catalysis. A growing number of enzymes are known to require a posttranslationally generated free radical for their proper functioning. The radical is generally used to remove a hydrogen atom from an unreactive position in the substrate, activating the substrate to undergo difficult chemistry. Some radical enzymes have unexpectedly been found to harbour a stable radical on an amino acid side‐chain that is distinct from the side‐chains of the active site region; during catalysis, the stable free radical interacts with the active site via radical transfer, and between turnovers it serves as a radical sink. Key Concepts Radical enzymes utilise single‐electron chemistry to enable difficult chemical reactions. Tyrosyl and tryptophanyl radicals are involved in a large number of biochemical reactions as initiators of catalysis or as partners in radical transfer pathways. Glycyl radicals are oxygen‐sensitive and have only been found in anaerobically expressed enzymes. Transient cysteinyl radicals are important intermediates in a number of radical enzymes. A transient 5′‐deoxyadensyl radical can be formed either by reductive cleavage of the S‐ adenosylmethionine cofactor in a radical SAM enzyme or by homolytic cleavage of the vitamin B12 coenzyme AdoCbl. All members of the ribonucleotide reductase enzyme family utilise a transient cysteinyl radical generated by a stable radical in a separate subunit (class I), cleavage of AdoCbl (class II), or a stable glycyl radical (class III).

show abstract

Structures of benzylsuccinate synthase elucidate roles of accessory subunits in glycyl radical enzyme activation and activity

Cited by 57 publications

References 33 publications

1,2-Propanediol Dehydration in Roseburia inulinivorans

1,2-Propanediol Dehydration in Roseburia inulinivorans

Substrate-bound Structures of Benzylsuccinate Synthase Reveal How Toluene Is Activated in Anaerobic Hydrocarbon Degradation

Radical Enzymes

Contact Info

Product

Resources

About