2.50
Hdl Handle:
http://hdl.handle.net/10541/108936
Title:
Biological and structural properties of MIP-1 alpha expressed in yeast.
Authors:
Clements, J M; Craig, S; Gearing, A J; Hunter, M G; Heyworth, Clare M; Dexter, T Michael; Lord, Brian I
Abstract:
The murine macrophage inflammatory proteins-1 alpha (MIP-1 alpha) and MIP-1 beta are distinct but closely related cytokines. Partially purified mixtures of the two proteins affect neutrophil function and cause local inflammation and fever. The particular properties of MIP-1 alpha have not been well studied, although it has been identified as being identical to an inhibitor of haemopoietic stem cell growth. We have expressed MIP-1 alpha in yeast cells and purified it to sequence homogeneity. Structural analysis of this biologically active material by circular dichroism and fluorescence spectroscopy confirms that MIP-1 alpha has a very similar secondary and tertiary structure to platelet factor 4 and interleukin 8 with which it shares limited sequence homology. The in-vitro stem cell inhibitory properties have been confirmed using a range of murine progenitor cells including purified bone marrow progenitor cells (FACS-1), the FDCP-mix A4 cell line, and spleen colony forming unit (CFU-S) populations. Plateau levels of inhibition of stem cell growth were achieved using concentrations of 0.15 micrograms/ml MIP-1 alpha. We have also demonstrated that MIP-1 alpha is active in vivo: 5 micrograms of MIP-1 alpha per mouse given as a bolus injection, protects stem cells from subsequent in-vitro killing by tritiated thymidine. MIP-1 alpha was also shown to enhance the proliferation of more committed progenitor granulocyte macrophage-colony forming cells (GM-CFC) in response to granulocyte macrophage-colony stimulating factor (GM-CSF).
Affiliation:
British Bio-technology, Cowley, Oxford.
Citation:
Biological and structural properties of MIP-1 alpha expressed in yeast. 1992, 4 (1):76-82 Cytokine
Journal:
Cytokine
Issue Date:
Jan-1992
URI:
http://hdl.handle.net/10541/108936
DOI:
10.1016/1043-4666(92)90040-X
PubMed ID:
1617159
Type:
Article
Language:
en
ISSN:
1043-4666
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorClements, J Men
dc.contributor.authorCraig, Sen
dc.contributor.authorGearing, A Jen
dc.contributor.authorHunter, M Gen
dc.contributor.authorHeyworth, Clare Men
dc.contributor.authorDexter, T Michaelen
dc.contributor.authorLord, Brian Ien
dc.date.accessioned2010-08-03T11:12:46Z-
dc.date.available2010-08-03T11:12:46Z-
dc.date.issued1992-01-
dc.identifier.citationBiological and structural properties of MIP-1 alpha expressed in yeast. 1992, 4 (1):76-82 Cytokineen
dc.identifier.issn1043-4666-
dc.identifier.pmid1617159-
dc.identifier.doi10.1016/1043-4666(92)90040-X-
dc.identifier.urihttp://hdl.handle.net/10541/108936-
dc.description.abstractThe murine macrophage inflammatory proteins-1 alpha (MIP-1 alpha) and MIP-1 beta are distinct but closely related cytokines. Partially purified mixtures of the two proteins affect neutrophil function and cause local inflammation and fever. The particular properties of MIP-1 alpha have not been well studied, although it has been identified as being identical to an inhibitor of haemopoietic stem cell growth. We have expressed MIP-1 alpha in yeast cells and purified it to sequence homogeneity. Structural analysis of this biologically active material by circular dichroism and fluorescence spectroscopy confirms that MIP-1 alpha has a very similar secondary and tertiary structure to platelet factor 4 and interleukin 8 with which it shares limited sequence homology. The in-vitro stem cell inhibitory properties have been confirmed using a range of murine progenitor cells including purified bone marrow progenitor cells (FACS-1), the FDCP-mix A4 cell line, and spleen colony forming unit (CFU-S) populations. Plateau levels of inhibition of stem cell growth were achieved using concentrations of 0.15 micrograms/ml MIP-1 alpha. We have also demonstrated that MIP-1 alpha is active in vivo: 5 micrograms of MIP-1 alpha per mouse given as a bolus injection, protects stem cells from subsequent in-vitro killing by tritiated thymidine. MIP-1 alpha was also shown to enhance the proliferation of more committed progenitor granulocyte macrophage-colony forming cells (GM-CFC) in response to granulocyte macrophage-colony stimulating factor (GM-CSF).en
dc.language.isoenen
dc.subjectHaematopoietic Stem Cellsen
dc.subject.meshAnimals-
dc.subject.meshBase Sequence-
dc.subject.meshCell Division-
dc.subject.meshCells, Cultured-
dc.subject.meshChemokine CCL4-
dc.subject.meshCircular Dichroism-
dc.subject.meshColony-Forming Units Assay-
dc.subject.meshCytokines-
dc.subject.meshDrug Synergism-
dc.subject.meshGenetic Vectors-
dc.subject.meshGranulocyte-Macrophage Colony-Stimulating Factor-
dc.subject.meshHematopoietic Stem Cells-
dc.subject.meshMacrophage Inflammatory Proteins-
dc.subject.meshMice-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshMonokines-
dc.subject.meshProtein Conformation-
dc.subject.meshRecombinant Fusion Proteins-
dc.subject.meshSaccharomyces cerevisiae-
dc.titleBiological and structural properties of MIP-1 alpha expressed in yeast.en
dc.typeArticleen
dc.contributor.departmentBritish Bio-technology, Cowley, Oxford.en
dc.identifier.journalCytokineen

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