Localization of Proteins to the 1,2-Propanediol Utilization Microcompartment by Non-native Signal Sequences Is Mediated by a Common Hydrophobic Motif

Jakobson, Christopher M.; Kim, Edward Y.; Slininger, Marilyn F.; Chien, Alex; Tullman‐Ercek, Danielle

doi:10.1074/jbc.m115.651919

Cited by 55 publications

(83 citation statements)

References 45 publications

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“…Studies indicate that a single N-terminal enzyme-targeting sequence can bind multiple shell proteins (38,39) and vice versa, i.e., that a single shell protein can bind multiple core enzymes through targeting sequence associations (59,62,63). Recent findings have also shown that targeting sequences from diverse MCP systems (Eut and Grp) can direct the encapsulation of proteins into the Pdu MCP via a conserved hydrophobic motif (36). Thus, the seemingly low specificity observed for core-shell interactions raises the question of whether MCP cores and shells (each of which are composed of 4 to 10 different proteins) have defined relative molecular orientations.…”

Section: Discussionmentioning

confidence: 99%

“…This idea developed from studies that showed the N-terminal 18 amino acids of the PduP core enzyme function as a targeting sequence that is necessary and sufficient for encapsulation of proteins within the Pdu MCP, as well as an extensive bioinformatic analysis that predicted that analogous short extensions are widely used in MCP assembly (35). Further work showed that short N-terminal extensions are used to target enzymes to diverse types of MCPs, both native and recombinant (27,33,34,36,39,(60)(61)(62). Mechanistic studies found that the N-terminal targeting sequences of PduP form an ␣-helix that binds to a short C-terminal helix of the PduA, PduJ, and/or PduK shell proteins to mediate encapsulation (38,39).…”

Section: Discussionmentioning

confidence: 99%

“…Binding occurs along conserved surfaces and mediates enzyme encapsulation (36,38,60,62). Site-directed mutagenesis of PduB ␣-helix at residues 6 to 18 indicated that a conserved hydrophobic patch formed by residues V7, I10, M11, and V14 was crucial to MCP assembly.…”

Section: Discussionmentioning

confidence: 99%

“…Spatial organization of enzymes by encapsulation within protein shells (as exemplified in MCP systems) has the ability to accelerate catalysis, prevent side reactions, and minimize the harmful effects of toxic intermediates. A key advance in this field has been the production of empty MCP shells which were then filled with heterologous cargo by fusing targeting/ encapsulation sequences to desired enzymes/proteins (33,36,(60)(61)(62)(64)(65)(66)(67)(68). Further development of this approach for efficient encapsulation of multiple enzymes with defined stoichiometries will likely require the use of various types of targeting sequences, as well as parameterization of the molecular interactions that define encapsulation.…”

Section: Discussionmentioning

confidence: 99%

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The N Terminus of the PduB Protein Binds the Protein Shell of the Pdu Microcompartment to Its Enzymatic Core

2017

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Bacterial microcompartments (MCPs) are extremely large proteinaceous organelles that consist of an enzymatic core encapsulated within a complex protein shell. A key question in MCP biology is the nature of the interactions that guide the assembly of thousands of protein subunits into a well-ordered metabolic compartment. In this report, we show that the N-terminal 37 amino acids of the PduB protein have a critical role in binding the shell of the 1,2-propanediol utilization (Pdu) microcompartment to its enzymatic core. Several mutations were constructed that deleted short regions of the N terminus of PduB. Growth tests indicated that three of these deletions were impaired MCP assembly. Attempts to purify MCPs from these mutants, followed by gel electrophoresis and enzyme assays, indicated that the protein complexes isolated consisted of MCP shells depleted of core enzymes. Electron microscopy substantiated these findings by identifying apparently empty MCP shells but not intact MCPs. Analyses of 13 site-directed mutants indicated that the key region of the N terminus of PduB required for MCP assembly is a putative helix spanning residues 6 to 18. Considering the findings presented here together with prior work, we propose a new model for MCP assembly.IMPORTANCE Bacterial microcompartments consist of metabolic enzymes encapsulated within a protein shell and are widely used to optimize metabolic process. Here, we show that the N-terminal 37 amino acids of the PduB shell protein are essential for assembly of the 1,2-propanediol utilization microcompartment. The results indicate that it plays a key role in binding the outer shell to the enzymatic core. We propose that this interaction might be used to define the relative orientation of the shell with respect to the core. This finding is of fundamental importance to our understanding of microcompartment assembly and may have application to engineering microcompartments as nanobioreactors for chemical production.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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The N Terminus of the PduB Protein Binds the Protein Shell of the Pdu Microcompartment to Its Enzymatic Core

2017

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“…For example, encapsulation efficiency can be improved by delayed expression of the cargo relative to the shell proteins 136 and by altering the primary structure of encapsulation peptides 137 . To identify the primary structure features that are important for encapsulation efficiency, encapsulation peptides were designed de novo 134 and reporter proteins that were fused to encapsulation peptides from GRM and EUT BMCs were encapsulated in a PDU BMC 138 , demonstrating the potential for using encapsulation peptides to attach proteins to diverse BMC shells.…”

Section: Bacterial Microcompartments In Bioengineeringmentioning

confidence: 99%

Bacterial microcompartments

et al. 2018

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Bacterial microcompartments (BMCs) are self-assembling organelles that consist of an enzymatic core that is encapsulated by a selectively permeable protein shell. The potential to form BMCs is widespread, found across the Kingdom Bacteria. BMCs have crucial roles in carbon dioxide fixation in autotrophs and the catabolism of organic substrates in heterotrophs. They contribute to the metabolic versatility of bacteria, providing a competitive advantage in specific environmental niches. Although BMCs were first visualized more than sixty years ago, it is mainly in the last decade that progress has been made in understanding their metabolic diversity and the structural basis of their assembly and function. This progress has not only heightened our understanding of their role in microbial metabolism but it is also beginning to enable their use in a variety of applications in synthetic biology. In this Review, we focus on recent insights into the structure, assembly, diversity and function of BMCs.

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Microbial Fermentation of Polyethylene Terephthalate (PET) Plastic Waste for the Production of Chemicals or Electricity**

2022

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Ideonella sakaiensis (I. sakaiensis) can grow on polyethylene terephthalate (PET) as the major carbon and energy source. Previous work has shown that PET conversion in the presence of oxygen released carbon dioxide and water while yielding adenosine triphosphate (ATP) through oxidative phosphorylation. This study demonstrates that I. sakaiensis is a facultative anaerobe that ferments PET to the feedstock chemicals acetate and ethanol in the absence of oxygen. In addition to PET, the pure monomer ethylene glycol (EG), the intermediate product ethanol, and the carbohydrate fermentation test substance maltose can also serve as fermenting substrates. Co-culturing of I. sakaiensis with the electrogenic and acetate-consuming Geobacter sulfurreducens produced electricity from PET or EG. This newly identified plastic fermentation process by I. sakaiensis provides thus a novel biosynthetic route to produce high-value chemicals or electricity from plastic waste streams.

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Localization of Proteins to the 1,2-Propanediol Utilization Microcompartment by Non-native Signal Sequences Is Mediated by a Common Hydrophobic Motif

Cited by 55 publications

References 45 publications

The N Terminus of the PduB Protein Binds the Protein Shell of the Pdu Microcompartment to Its Enzymatic Core

The N Terminus of the PduB Protein Binds the Protein Shell of the Pdu Microcompartment to Its Enzymatic Core

Bacterial microcompartments

Microbial Fermentation of Polyethylene Terephthalate (PET) Plastic Waste for the Production of Chemicals or Electricity**

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