PMID- 10347159
OWN - NLM
STAT- MEDLINE
DCOM- 19990701
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 23
DP  - 1999 Jun 4
TI  - Identification of amino acid residues critical for aggregation of human CC
      chemokines macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and RANTES.
      Characterization of active disaggregated chemokine variants.
PG  - 16077-84
AB  - Human CC chemokines macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and 
      RANTES (regulated on activation normal T cell expressed) self-associate to form
      high-molecular mass aggregates. To explore the biological significance of
      chemokine aggregation, nonaggregating variants were sought. The phenotypes of 105
      hMIP-1alpha variants generated by systematic mutagenesis and expression in yeast 
      were determined. hMIP-1alpha residues Asp26 and Glu66 were critical to the
      self-association process. Substitution at either residue resulted in the
      formation of essentially homogenous tetramers at 0.5 mg/ml. Substitution of
      identical or analogous residues in homologous positions in both hMIP-1beta and
      RANTES demonstrated that they were also critical to aggregation. Our analysis
      suggests that a single charged residue at either position 26 or 66 is
      insufficient to support extensive aggregation and that two charged residues must 
      be present. Solution of the three-dimensional NMR structure of hMIP-1alpha has
      enabled comparison of these residues in hMIP-1beta and RANTES. Aggregated and
      disaggregated forms of hMIP-1alpha, hMIP-1beta, and RANTES generally have
      equivalent G-protein-coupled receptor-mediated biological potencies. We have
      therefore generated novel reagents to evaluate the role of hMIP-1alpha,
      hMIP-1beta, and RANTES aggregation in vitro and in vivo. The disaggregated
      chemokines retained their human immunodeficiency virus (HIV) inhibitory
      activities. Surprisingly, high concentrations of RANTES, but not disaggregated
      RANTES variants, enhanced infection of cells by both M- and T-tropic HIV
      isolates/strains. This observation has important implications for potential
      therapeutic uses of chemokines implying that disaggregated forms may be necessary
      for safe clinical investigation.
FAU - Czaplewski, L G
AU  - Czaplewski LG
AD  - British Biotech Pharmaceuticals Ltd., Watlington Road, Oxford OX4 5LY, United
      Kingdom. czaplewski@britbio.co.uk
FAU - McKeating, J
AU  - McKeating J
FAU - Craven, C J
AU  - Craven CJ
FAU - Higgins, L D
AU  - Higgins LD
FAU - Appay, V
AU  - Appay V
FAU - Brown, A
AU  - Brown A
FAU - Dudgeon, T
AU  - Dudgeon T
FAU - Howard, L A
AU  - Howard LA
FAU - Meyers, T
AU  - Meyers T
FAU - Owen, J
AU  - Owen J
FAU - Palan, S R
AU  - Palan SR
FAU - Tan, P
AU  - Tan P
FAU - Wilson, G
AU  - Wilson G
FAU - Woods, N R
AU  - Woods NR
FAU - Heyworth, C M
AU  - Heyworth CM
FAU - Lord, B I
AU  - Lord BI
FAU - Brotherton, D
AU  - Brotherton D
FAU - Christison, R
AU  - Christison R
FAU - Craig, S
AU  - Craig S
FAU - Cribbes, S
AU  - Cribbes S
FAU - Edwards, R M
AU  - Edwards RM
FAU - Evans, S J
AU  - Evans SJ
FAU - Gilbert, R
AU  - Gilbert R
FAU - Morgan, P
AU  - Morgan P
FAU - Randle, E
AU  - Randle E
FAU - Schofield, N
AU  - Schofield N
FAU - Varley, P G
AU  - Varley PG
FAU - Fisher, J
AU  - Fisher J
FAU - Waltho, J P
AU  - Waltho JP
FAU - Hunter, M G
AU  - Hunter MG
LA  - eng
SI  - PDB/1B50
SI  - PDB/1B53
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Amino Acids)
RN  - 0 (Chemokine CCL3)
RN  - 0 (Chemokine CCL4)
RN  - 0 (Chemokine CCL5)
RN  - 0 (Macrophage Inflammatory Proteins)
RN  - 0 (Peptide Library)
SB  - IM
MH  - Amino Acid Sequence
MH  - Amino Acids/*analysis
MH  - Cell Line
MH  - Chemokine CCL3
MH  - Chemokine CCL4
MH  - Chemokine CCL5/*chemistry/genetics
MH  - HIV Infections/metabolism
MH  - HIV-1
MH  - Humans
MH  - Macrophage Inflammatory Proteins/*chemistry/genetics
MH  - Magnetic Resonance Spectroscopy
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Peptide Library
MH  - Protein Conformation
MH  - Structure-Activity Relationship
EDAT- 1999/05/29 00:00
MHDA- 1999/05/29 00:01
CRDT- 1999/05/29 00:00
PHST- 1999/05/29 00:00 [pubmed]
PHST- 1999/05/29 00:01 [medline]
PHST- 1999/05/29 00:00 [entrez]
AID - 10.1074/jbc.274.23.16077 [doi]
AID - S0021-9258(19)72944-5 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 Jun 4;274(23):16077-84. doi: 10.1074/jbc.274.23.16077.