Hostname: page-component-76fb5796d-25wd4 Total loading time: 0 Render date: 2024-04-26T18:08:15.025Z Has data issue: false hasContentIssue false
  • English
  • Français

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 ,
T. MEYERS ,
D. LOWNE ,
P.G. VARLEY ,
J.R.P. ARNOLD ,
P. TAN ,
J.-C. YANG ,
L.G. CZAPLEWSKI ,
T. DUDGEON  and
J. FISHER
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
Get access

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A sequence variant of human MIP-1α, in which Asp26 has been replaced by Ala, has been chemically modified by the addition of 13C-labeled methyl groups at each of the lysine residues and the N-terminus.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.

Keywords

13C NMRchemical modification of lysineshuman MIP-1α
Type
FOR THE RECORD
Information
Protein Science , Volume 9 , Issue 10 , October 2000 , pp. 2047 - 2053
Copyright
© 2000 The Protein Society

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)