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PMID: 21821773 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

The N-terminal region of the medium subunit (PduD) packages adenosylcobalamin-dependent diol dehydratase (PduCDE) into the Pdu microcompartment.

Journal of bacteriology ·Vol. 193 ·No. 20 ·2011-10-00 ·Pages 5623-8

Fan C, Bobik TA

Abstract

Salmonella enterica produces a proteinaceous microcompartment for B(12)-dependent 1,2-propanediol utilization (Pdu MCP). The Pdu MCP consists of catabolic enzymes encased within a protein shell, and its function is to sequester propionaldehyde, a toxic intermediate of 1,2-propanediol degradation. We report here that a short N-terminal region of the medium subunit (PduD) is required for packaging the coenzyme B(12)-dependent diol dehydratase (PduCDE) into the lumen of the Pdu MCP. Analysis of soluble cell extracts and purified MCPs by Western blotting showed that the PduD subunit mediated packaging of itself and other subunits of diol dehydratase (PduC and PduE) into the Pdu MCP. Deletion of 35 amino acids from the N terminus of PduD significantly impaired the packaging of PduCDE with minimal effects on its enzyme activity. Western blotting showed that fusing the 18 N-terminal amino acids of PduD to green fluorescent protein or glutathione S-transferase resulted in the association of these fusion proteins with the MCP. Immunoprecipitation tests indicated that the fusion proteins were encapsulated inside the MCP shell.

MeSH Terms
Bacterial Proteins/chemistry,genetics,metabolism Cobamides/metabolism Cytoplasmic Granules/chemistry,enzymology,genetics Molecular Sequence Data Propanediol Dehydratase/chemistry,genetics,metabolism Propylene Glycol/metabolism Protein Structure, Tertiary Protein Subunits/chemistry,genetics,metabolism Salmonella enterica/chemistry,enzymology,genetics
Chemicals
Bacterial Proteins Cobamides Protein Subunits Propylene Glycol Propanediol Dehydratase cobamamide
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Fan Chenguang
Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011, USA.
Bobik Thomas A
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
1098-5530
DOI
10.1128/JB.05661-11
Published
2011-10-00
Epub
2011-00-05
Pages
5623-8
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC3187188
Subset
IM
Grants
NIAID NIH HHS · R01 AI081146 · United States
NIAID NIH HHS · AI081146 · United States
External Links
PubMed source DOI: 10.1128/JB.05661-11 PMC Full Text View journal details
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