Analysis of Chaperone Function in Vitro

Büchner, Johannes; Walter, Stefan

doi:10.1002/9783527619498.ch38

Cited by 6 publications

(3 citation statements)

References 150 publications

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“…The column was washed with 100 mM imidazole and recombinant protein eluted with 500 mM imidazole/6 M urea. The eluate was concentrated to 20 mg/ml using 3 kDa 15 ml Amicon Ultra centrifuge concentration devices (Millipore) and refolded by drop wise addition to a 20-fold greater volume of a range of refolding buffers [52]. The soluble material was buffer-exchanged into PBS using a PD10 column (GE).…”

Section: Methodsmentioning

confidence: 99%

A Granulin-Like Growth Factor Secreted by the Carcinogenic Liver Fluke, Opisthorchis viverrini, Promotes Proliferation of Host Cells

et al. 2009

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The human liver fluke, Opisthorchis viverrini, infects millions of people throughout south-east Asia and is a major cause of cholangiocarcinoma, or cancer of the bile ducts. The mechanisms by which chronic infection with O. viverrini results in cholangiocarcinogenesis are multi-factorial, but one such mechanism is the secretion of parasite proteins with mitogenic properties into the bile ducts, driving cell proliferation and creating a tumorigenic environment. Using a proteomic approach, we identified a homologue of human granulin, a potent growth factor involved in cell proliferation and wound healing, in the excretory/secretory (ES) products of the parasite. O. viverrini granulin, termed Ov-GRN-1, was expressed in most parasite tissues, particularly the gut and tegument. Furthermore, Ov-GRN-1 was detected in situ on the surface of biliary epithelial cells of hamsters experimentally infected with O. viverrini. Recombinant Ov-GRN-1 was expressed in E. coli and refolded from inclusion bodies. Refolded protein stimulated proliferation of murine fibroblasts at nanomolar concentrations, and proliferation was inhibited by the MAPK kinase inhibitor, U0126. Antibodies raised to recombinant Ov-GRN-1 inhibited the ability of O. viverrini ES products to induce proliferation of murine fibroblasts and a human cholangiocarcinoma cell line in vitro, indicating that Ov-GRN-1 is the major growth factor present in O. viverrini ES products. This is the first report of a secreted growth factor from a parasitic worm that induces proliferation of host cells, and supports a role for this fluke protein in establishment of a tumorigenic environment that may ultimately manifest as cholangiocarcinoma.

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Section: Methodsmentioning

confidence: 99%

A Granulin-Like Growth Factor Secreted by the Carcinogenic Liver Fluke, Opisthorchis viverrini, Promotes Proliferation of Host Cells

et al. 2009

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“…(iii) Molecular chaperones prevent protein aggregation by interacting with hydrophobic patches exposed by the folding substrate. By definition, molecular chaperones are a family of cellular proteins that assist non-covalent folding and unfolding, as well as assembly or disassembly of their substrates while they are not part of the final structure [22]. Thereby, they help in blocking alternative assembly or folding pathways that produce non-functional structures, e.g., by self-aggregation.…”

Section: Introductionmentioning

confidence: 99%

Real-time protein NMR spectroscopy and investigation of assisted protein folding

Kumar

Balbach

2015

Biochimica et Biophysica Acta (BBA) - General Subjects

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“… Hüttemann and Love observe that there is an ongoing debate concerning the nature of the causal contribution of chaperones to folding. According toEllis (1998) for instance, chaperones provide additional "steric information", whileBuchner & Walter (2005) deny this. The ethos of their narrative implies that Hüttemann and Love side with Ellis (1998), even though the reasons are unclear.…”

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confidence: 99%

A relational-constructionist account of protein macrostructure and function

2020

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One of the foundational problems of biochemistry concerns the conceptualisation of the relationship between the composition, structure and function of macromolecules like proteins. Part of the recent philosophical literature displays a reductionist bias, that is, the endorsement of a form of microstructuralism mirroring an out-dated biochemical conceptualisation. We shall argue that such microstructuralist approaches are ultimately committed to a potentialist form of micro-predeterminism whereby the macrostructure and function of proteins is accounted for solely in terms of the intrinsic properties and potentialities of the components of the primary structure as if they were selfcontained or essentially immutable entities. We shall instead suggest that a conceptualisation of the relationship between proteins' composition, structure and function consistent with contemporary biochemical practice should account also for the causal role of the cellular, organismal and environmental relations in protein development. The analysis of the folding process we propose suggests that microstructure-laden reductionist approaches are ontologically indefensible. Rather than a potentialist form of micro-predeterminism, our

show abstract

Analysis of Chaperone Function in Vitro

Cited by 6 publications

References 150 publications

A Granulin-Like Growth Factor Secreted by the Carcinogenic Liver Fluke, Opisthorchis viverrini, Promotes Proliferation of Host Cells

A Granulin-Like Growth Factor Secreted by the Carcinogenic Liver Fluke, Opisthorchis viverrini, Promotes Proliferation of Host Cells

Real-time protein NMR spectroscopy and investigation of assisted protein folding

A relational-constructionist account of protein macrostructure and function

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