Characterization of the quaternary structure and conformational properties of the human stem cell inhibitor protein LD78 in solution.
The human LD78 protein (sometimes referred to as human macrophage inflammatory protein-1 alpha) has been shown to protect multipotential hemopoietic stem cells from the effects of cytotoxic agents. Administration of the recombinant stem cell inhibitor molecule LD78 as an adjunct to chemotherapy has potential clinical benefit in reducing or preventing the neutropenia associated with this treatment. At physiological ionic strength, the 8-kDa LD78 molecule exists as soluble, heterogeneous, multimeric complexes of mass ranging from 100 to > 250 kDa. The hydrodynamic and structural properties of LD78 have been determined in various buffer solutions using analytical ultracentrifugation, circular dichroism, and fluorescence spectroscopy. The results demonstrate that defined, homogeneous monomer and tetramer forms of LD78 can be prepared which display distinct conformational properties. The combined use of hydrodynamic and spectroscopic analysis provides an insight into the pathway and molecular mechanics of LD78 self-association.
British Bio-technology Ltd., Cowley, Oxford, U.K.
S R
SR
Patel
S
S
Evans
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K
Dunne
G C
GC
Knight
P J
PJ
Morgan
P G
PG
Varley
S
S
Craig
D000085:Acetates; D019342:Acetic Acid; D000097:Acetonitriles; D054407:Chemokine CCL4; D002942:Circular Dichroism; D016207:Cytokines / Q000737:chemistry*; D004563:Electrochemistry; D005454:Fluorescence Polarization; D006801:Humans; D006863:Hydrogen-Ion Concentration; D046911:Macromolecular Substances; D019402:Macrophage Inflammatory Proteins; D008970:Molecular Weight; D015846:Monokines / Q000737:chemistry*; D011487:Protein Conformation; D011994:Recombinant Proteins; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017421:Sequence Analysis; D012996:Solutions; D013050:Spectrometry, Fluorescence
D000085:Acetates; D000097:Acetonitriles; D054407:Chemokine CCL4; D016207:Cytokines; D046911:Macromolecular Substances; D019402:Macrophage Inflammatory Proteins; D015846:Monokines; D011994:Recombinant Proteins; D012996:Solutions; D019342:Acetic Acid; C032159:acetonitrile