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PMID: 20308536 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S.

Short N-terminal sequences package proteins into bacterial microcompartments.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 107 ·No. 16 ·2010-04-20 ·Pages 7509-14

Fan C, Cheng S, Liu Y, Escobar CM, Crowley CS, Jefferson RE, Yeates TO, Bobik TA

Abstract

Hundreds of bacterial species produce proteinaceous microcompartments (MCPs) that act as simple organelles by confining the enzymes of metabolic pathways that have toxic or volatile intermediates. A fundamental unanswered question about bacterial MCPs is how enzymes are packaged within the protein shell that forms their outer surface. Here, we report that a short N-terminal peptide is necessary and sufficient for packaging enzymes into the lumen of an MCP involved in B(12)-dependent 1,2-propanediol utilization (Pdu MCP). Deletion of 10 or 14 amino acids from the N terminus of the propionaldehyde dehydrogenase (PduP) enzyme, which is normally found within the Pdu MCP, substantially impaired packaging, with minimal effects on its enzymatic activity. Fusion of the 18 N-terminal amino acids from PduP to GFP, GST, or maltose-binding protein resulted in their encapsulation within MCPs. Bioinformatic analyses revealed N-terminal extensions in two additional Pdu proteins and three proteins from two unrelated MCPs, suggesting that N-terminal peptides may be used to package proteins into diverse MCPs. The potential uses of MCP assembly principles in nature and in biotechnology are discussed.

MeSH Terms
Amino Acid Sequence Amino Acids/chemistry Bacteria/metabolism Computational Biology/methods Green Fluorescent Proteins/chemistry Maltose-Binding Proteins Microscopy, Fluorescence/methods Models, Genetic Molecular Sequence Data Periplasmic Binding Proteins/chemistry Propylene Glycol/chemistry Protein Structure, Tertiary Recombinant Fusion Proteins/chemistry Salmonella enterica/metabolism Sequence Homology, Amino Acid Vitamin B 12/metabolism
Chemicals
Amino Acids Maltose-Binding Proteins Periplasmic Binding Proteins Recombinant Fusion Proteins Green Fluorescent Proteins Propylene Glycol Vitamin B 12
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Fan Chenguang
Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, Ames, IA 50011, USA.
Cheng Shouqiang
Liu Yu
Escobar Cristina M
Crowley Christopher S
Jefferson Robert E
Yeates Todd O
Bobik Thomas A
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
1091-6490
DOI
10.1073/pnas.0913199107
Published
2010-04-20
Epub
2010-00-22
Pages
7509-14
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC2867708
Subset
IM
Grants
NIAID NIH HHS · R01 AI081146 · United States
NIAID NIH HHS · AI081146 · United States
Corrections
CommentIn
PMID: 20400692
External Links
PubMed source DOI: 10.1073/pnas.0913199107 PMC Full Text View journal details
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