Purification and characterization of soluble forms of human and rat stem cell factor recombinantly expressed by Escherichia coli and by Chinese hamster ovary cells

Langley, Keith; Wypych, Jette; Mendiaz, Elizabeth A.; Clogston, Christi L.; Parker, Vann P.; Farrar, D; Brothers, Mark O.; Satygal, Vasuki N.; Leslie, I.; Birkett, Neal C.; Smith, Kent; Baltera, Robert F.; Lyons, Daniel F.; Hogan, Jeffery M.; Crandall, Craig A.; Boone, Thomas C.; Pope, Joseph A.; Karkare, Subhash B.; Zsebo, Krisztina M.; Sachdev, Raj; Lu, Hsieng S.

doi:10.1016/0003-9861(92)90482-c

Cited by 54 publications

(31 citation statements)

References 33 publications

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“…Inclusion body formation, as might be seen in the insoluble fraction of the cell lysate, is hardly observed. Up to the present, SCF expression in E. coli has always led to insoluble inclusion bodies (Langley et al, 1992;Lili et al, 2006;Potala and Verma, 2010;Su et al, 2006;Wang et al, 2008), thus the formation of soluble TRX-mSCF is obviously due to the TRX presence.…”

Section: Expression Of Trx-mscfmentioning

confidence: 86%

“…The soluble protein is heavily N-and O-glycosylated, has extensive secondary structures and is dimeric (non covalently associated) under non-denaturating conditions (Arakawa et al, 1991;Langley et al, 1992;Zhang et al, 2000). The presence or absence of the carbohydrate moieties does not influence the biological activity (Flanagan and Leder, 1990;Zsebo et al, 1990), whereas the two intramolecular disulfide bonds of SCF are essential (Jones et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

“…Human and rat SCF were expressed in E. coli, in COS-7 cells and in CHO cells (Arakawa et al, 1991;Flanagan and Leder, 1990;Langley et al, 1992;Martin et al, 1990). The simultaneous renaturation and purification of E. coli-derived human SCF has been described as well (Lili et al, 2006;Wang et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

“…To date, SCF has exclusively been expressed in E. coli as insoluble and inactive aggregates (Arakawa et al, 1991;Langley et al, 1994;Langley et al, 1992;Lili et al, 2006;Martin et al, 1990;Wang et al, 2008;Wang et al, 2004). The current strategy for recovery of active SCF from inclusion bodies includes solubilization, oxidation and refolding steps (Wang et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Expression and purification of bioactive soluble murine stem cell factor from recombinant Escherichia coli using thioredoxin as fusion partner

Bals

Schambach

Meyer

et al. 2011

Journal of Biotechnology

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Section: Expression Of Trx-mscfmentioning

confidence: 86%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Expression and purification of bioactive soluble murine stem cell factor from recombinant Escherichia coli using thioredoxin as fusion partner

Bals

Schambach

Meyer

et al. 2011

Journal of Biotechnology

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“…Whereas soluble Kitl forms non-covalent homodimers, each Kitl monomer contains two intra-chain disulfide bonds, C4-C89 and C43-C138 (Arakawa et al, 1991;Jiang et al, 2000;Langley et al, 1992;Zhang et al, 2000). The Kitl sequences represent the functional core of Kitl that contains the dimer interface and the domain involved in Kit receptor binding and are sufficient to mediate cell proliferation activity (Langley et al, 1994).…”

Section: Evaluation Of the Effect Of Deletions On Kitl Sheddingmentioning

confidence: 99%

Different ADAMs have distinct influences on Kit ligand processing: phorbol-ester-stimulated ectodomain shedding of Kitl1 by ADAM17 is reduced by ADAM19

Kawaguchi

Horiuchi

Becherer

et al. 2007

Journal of Cell Science

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Kit ligand (Kitl), the ligand for the Kit receptor tyrosine kinase, plays important roles in hematopoiesis, gametogenesis and melanogenesis. Kitl is synthesized as a membrane-anchored precursor that can be processed to produce the soluble growth factor. Here, we evaluated the role of ADAM (a disintegrin and metalloprotease) metalloproteases in ectodomain shedding of Kitl. We found that both ADAM17 and ADAM19 affect Kitl1 shedding, albeit in different ways. Overexpression of ADAM19 resulted in decreased levels of Endo-H-resistant mature Kitl1, thereby reducing the amount of Kitl that is shed from cells following stimulation with phorbol esters. ADAM17 was identified as the major phorbol-ester-stimulated sheddase of Kitl1, whereas ADAMs 8, 9, 10, 12 and 15 were not required for this process. ADAM17 also emerged as the major constitutive and phorbol-ester-stimulated sheddase of Kitl2 in mouse embryonic fibroblasts. Mutagenesis of the juxtamembrane domain of Kitl2 showed no stringent sequence requirement for cleavage by ADAM17, although two nonadjacent stretches of four amino acid residues were identified that are required for Kitl2 shedding. Taken together, this study identifies a novel sheddase, ADAM17, for Kitl1 and Kitl2, and demonstrates that ADAM19 can reduce ADAM17-dependent phorbol-ester-stimulated Kitl1 ectodomain shedding.

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Computational simulations of stem-cell factor/c-Kit receptor interaction

1996

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Stem-cell factor (SCF) is a noncovalent homodimeric cytokine that exhibits profound biological function in the early stages of hematopoiesis by binding to a cell surface tyrosine kinase receptor that is encoded by the c-Kit proto-oncogene. The results obtained from a combined implementation of homology-based molecular modeling and computational simulations in the study of species-specific SCF/ c-Kit interactions are reported. The structural models of the human and rat SCF ligands are based on the close structural similarity to the cytokine M-CSF, whose C alpha structure has recently become available. The constant domains of the human Fc fragment are used as a template for the ligand binding domains of the c-Kit receptor. The factors responsible for the stabilization of the SCF quaternary structure and the molecular determinants for ligand recognition and ligand specificity have been identified by assessing the conformational, topographical, and dynamic features of the isolated ligands and of the ligand-receptor complexes.

show abstract

Purification and characterization of soluble forms of human and rat stem cell factor recombinantly expressed by Escherichia coli and by Chinese hamster ovary cells

Cited by 54 publications

References 33 publications

Expression and purification of bioactive soluble murine stem cell factor from recombinant Escherichia coli using thioredoxin as fusion partner

Expression and purification of bioactive soluble murine stem cell factor from recombinant Escherichia coli using thioredoxin as fusion partner

Different ADAMs have distinct influences on Kit ligand processing: phorbol-ester-stimulated ectodomain shedding of Kitl1 by ADAM17 is reduced by ADAM19

Computational simulations of stem-cell factor/c-Kit receptor interaction

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