The DOI matches its Crossref record; 42 of the 42 references it cites were found in a registry or an index.
Purification, Bioactivity, and Secondary Structure Analysis of Mouse and Human Macrophage Migration Inhibitory Factor (MIF)
Abstract
The cytokine macrophage migration inhibitory factor (MIF) has been identified to be secreted by the pituitary gland and the monocyte/macrophage and to play an important role in endotoxic shock. Despite the recent molecular cloning of a human T-cell MIF, characterization of the biochemical and biological properties of this protein has remained incomplete because substantial quantities of purified, recombinant, or native MIF have not been available. We describe the cloning of mouse MIF from anterior pituitary cells (AtT-20) and the purification of native MIF from mouse liver by sequential ion exchange and reverse-phase chromatography. For comparison purposes, human MIF was cloned from the Jurkat T-cell line and also characterized. Mouse and human MIF were highly homologous (90% identity over 115 amino acids). Recombinant mouse and human MIF were expressed in Escherichia coli and purified in milligram quantities by a simple two-step procedure. The molecular weight of native mouse MIF (12.5 kDa monomer) was identical with that of recombinant mouse MIF as assessed by gel electrophoresis and mass spectroscopy. No significant post-translational modifications were detected despite the presence of two potential N-linked glycosylation sites. Recombinant MIF inhibited monocyte migration in a dose-dependent fashion, and both recombinant and native MIF-exhibited comparable biological activities. MIF induced the secretion of tumor necrosis factor-alpha and stimulated nitric oxide production by macrophages primed with interferon-gamma. Circular dichroism spectroscopy revealed that bioactive mouse and human MIF exhibit a highly ordered, three-dimensional structure with a significant percentage of beta-sheet and alpha-helix conformation. Guanidine hydrochloride-induced unfolding experiments demonstrated that MIF is of low to moderate thermodynamic stability. These studies establish the biochemical identity of native and recombinant MIF and provide a first insight into the three-dimensional structural properties of this critical inflammatory mediator.
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Cites
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Cited by
Regulation of the Hypothalamic-Pituitary-Adrenal Axis by Cytokines: Actions and Mechanisms of Action
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MIF Signal Transduction Initiated by Binding to CD74
Leng, L., Metz, C. N., Fang, Y., Xu, J., Donnelly, S., Baugh, J., Delohery, T., Chen, Y., Mitchell, R. A., & Bucala, R. (2003) · The Journal of Experimental Medicine · doi:10.1084/jem.20030286 · Cited by 1,154
Self-citationAllergy Diagnostic Testing: An Updated Practice Parameter
Bernstein, I. L., Li, J. T., Bernstein, D. I., Hamilton, R., Spector, S. L., Tan, R., Sicherer, S., Golden, D. B. K., Khan, D. A., Nicklas, R. A., et al. (2008) · Annals of Allergy, Asthma & Immunology · doi:10.1016/s1081-1206(10)60305-5 · Cited by 723
CD44 Is the Signaling Component of the Macrophage Migration Inhibitory Factor-CD74 Receptor Complex
Shi, X., Leng, L., Wang, T., Wang, W., Du, X., Li, J., McDonald, C., Chen, Z., Murphy, J. W., Lolis, E., et al. (2006) · Immunity · doi:10.1016/j.immuni.2006.08.020 · Cited by 677
Self-citationMitchell, R. A., Liao, H., Chesney, J., Fingerle-Rowson, G., Baugh, J., David, J., & Bucala, R. (2002) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.012511599 · Cited by 622
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Metadata
- Authors
- Bernhagen, J., Mitchell, R. A., Calandra, T., Voelter, W., Cerami, A., & Bucala, R.
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1994-11-01
- Volume
- 33
- Issue
- 47
- Pages
- 14144-14155
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W1979244891
- Cited by
- 408
- References
- 42
- FWCI
- 3.87
- Type
- journal article
- Language
- en
- Open access
- closed
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- Bernhagen, J., Mitchell, R. A., Calandra, T., Voelter, W., Cerami, A., & Bucala, R. (1994). Purification, Bioactivity, and Secondary Structure Analysis of Mouse and Human Macrophage Migration Inhibitory Factor (MIF). Biochemistry. https://doi.org/10.1021/bi00251a025
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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