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Chemical modification of a variant of human MIP-1α; implications for dimer structure

Published online by Cambridge University Press:  15 December 2000

J.T. ASHFIELD
Affiliation:
School of Chemistry, University of Leeds, Leeds LS2 9JT, United Kingdom
T. MEYERS
Affiliation:
Current address: Cambridge Antibody Technology, The Science Park, Melbourn, Cambridgeshire SG8 6JJ, United Kingdom.
D. LOWNE
Affiliation:
British Biotech, Watlington Road, Cowley, Oxford OX4 5LY, United Kingdom
P.G. VARLEY
Affiliation:
Current address: Cambridge Antibody Technology, The Science Park, Melbourn, Cambridgeshire SG8 6JJ, United Kingdom.
J.R.P. ARNOLD
Affiliation:
School of Biology, University of Leeds, Leeds LS2 9JT, United Kingdom
P. TAN
Affiliation:
British Biotech, Watlington Road, Cowley, Oxford OX4 5LY, United Kingdom
J.-C. YANG
Affiliation:
MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom
L.G. CZAPLEWSKI
Affiliation:
British Biotech, Watlington Road, Cowley, Oxford OX4 5LY, United Kingdom
T. DUDGEON
Affiliation:
British Biotech, Watlington Road, Cowley, Oxford OX4 5LY, United Kingdom
J. FISHER
Affiliation:
School of Chemistry, University of Leeds, Leeds LS2 9JT, United Kingdom
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Abstract

A sequence variant of human MIP-1α, in which Asp26 has been replaced by Ala, has been chemically modified by the addition of 13C-labeled methyl groups at each of the lysine residues and the N-terminus. The sites of methylation have been verified by a combination of MALDI-TOF mass spectrometric experiments and tryptic digestion followed by N-terminal mapping. The effect of the modification on the structure and activity of the protein have been determined by analytical ultra-centrifugation, 13C NMR spectroscopy and receptor binding studies. The results of these experiments suggest that huMIP-1α D26A (BB10010), when present as a dimer, adopts a globular structure, like MCP-3, rather than the elongated or cylindrical structure determined for dimers of huMIP-1β and RANTES.

Type
FOR THE RECORD
Copyright
© 2000 The Protein Society

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