Using an E. coli Type 1 secretion system to secrete the mammalian, intracellular protein IFABP in its active form

Schwarz, Christian; Landsberg, Christine D.; Lenders, Michael H. H.; Smits, Sander H. J.; Schmitt, Lutz

doi:10.1016/j.jbiotec.2012.02.005

Cited by 26 publications

(26 citation statements)

References 42 publications

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“…Chimeric substrates have also been widely analyzed in particular for biotechnological or vaccinal purposes [99,[131][132][133][134]. These studies revealed (i) the importance of the RTX motifs for secretion, although this varies widely among the secretion apparatus and chimeric proteins tested [24,28,91,135]; (ii) a prerequisite for efficient secretion of the protein substrate is that it can unfold, likely to pass through the TolC channel, about 30 Å wide [115,116]; and (iii) premature folding of the protein substrates or stabilization of their 3D fold (e.g., via cytoplasmic disulfide bond formation) prevents secretion by T1SS [99,101,133,136]. Chaperones, like SecB or GroEL are not needed for HlyA or PtrB secretion [19,100], and it has been proposed that the RTX motifs could act as an internal chaperone that can prevent the folding of the protein substrate [17].…”

Section: Assembly Of the T1ss Apparatusmentioning

confidence: 99%

Structure and function of RTX toxins

Chenal

Sotomayor-Pérez

Ladant

2015

The Comprehensive Sourcebook of Bacterial Protein Toxins

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Section: Assembly Of the T1ss Apparatusmentioning

confidence: 99%

Structure and function of RTX toxins

Chenal

Sotomayor-Pérez

Ladant

2015

The Comprehensive Sourcebook of Bacterial Protein Toxins

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“…In favor of the activating model is the fact that many fusion proteins are secreted by the hemolysin T1SS (Mackman et al, 1987;Kenny et al, 1991;Blight and Holland, 1994;Gentschev et al, 2002;Bakkes et al, 2010;Schwarz et al, 2012). These proteins are fused at their C-terminus to a fragment of HlyA that contains the secretion signal.…”

Section: The Role Of the Secretion Signalmentioning

confidence: 99%

Molecular insights into type I secretion systems

Lenders¹,

Reimann²,

Smits

et al. 2013

Biological Chemistry

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Type 1 secretion systems are one of the main machineries in Gram-negative bacteria involved in the secretion of a wide range of substrates from the cytoplasm across the inner and outer membrane in one step to the extracellular space. The range of substrates varies from small proteins up to large surface layer proteins of about 900 kDa. Most of the substrates have a non-cleavable C-terminal secretion signal and so-called GG repeats that are able to bind calcium ions. The translocator complex is composed of a trimeric outer membrane protein that provides a pore in the outer membrane. A multimeric membrane fusion protein spans the periplasm and forms a continuous channel connecting the outer membrane protein with a dimeric ATP-binding cassette transporter in the inner membrane. The ATP-binding cassette-transporter is thought to form a channel through the inner membrane and energizes the transport process. This review will provide a detailed view of the components of the translocator and will summarize structural as well as functional data.

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“…9 Recently, two publications investigated these limitations. 8,13 Here, the maltose binding protein (MalE) and the intestinal fatty acid binding protein (IFABP) could initially not be secreted when fused to HlyAc. However, the use of slow-folding proteins with decreased folding kinetics allowed the secretion into the culture medium.…”

Section: Introductionmentioning

confidence: 99%

“…However, the use of slow-folding proteins with decreased folding kinetics allowed the secretion into the culture medium. 8,13 Interestingly, the mutant IFABP(G121V), accumulates in E. coli as inclusion bodies, whereas secreted IFABP(G121V) was soluble and active. Taken together, both studies suggest a rational to secrete proteins of interest in good yields and in a soluble, functional state with an E. coli T1SS.…”

Section: Introductionmentioning

confidence: 99%