PMID- 7531692
OWN - NLM
STAT- MEDLINE
DCOM- 19950316
LR  - 20210210
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 270
IP  - 6
DP  - 1995 Feb 10
TI  - Mutation of Leu25 and Val27 introduces CC chemokine activity into interleukin-8.
PG  - 2716-21
AB  - Interleukin-8 (IL-8) is a member of the CXC branch of the chemokine superfamily 
      and activates neutrophils but not monocytes. The related CC chemokine branch, 
      which includes monocyte chemoattractant protein-1 (MCP-1) and RANTES are potent 
      chemoattractants for monocytes but not neutrophils. Examination of the sequences 
      of the CXC chemokines reveals that the highly conserved leucine, corresponding to 
      Leu25 in IL-8, is always replaced by tyrosine in CC chemokines. There is also a 
      high degree of conservation among the CXC chemokines of the adjacent Val27 
      residue, which points out from the same side of the beta-sheet as Leu25. In 
      RANTES, Val27 is also replaced by a tyrosine. In order to investigate the role of 
      these residues in controlling cell specificity, we have made the single mutants 
      Leu25-->Tyr, Val27-->Tyr and the double mutant Leu25-->Tyr, Val27--> Tyr of IL-8. 
      These proteins have been expressed in Escherichia coli and purified to 
      homogeneity from inclusion body material. All three mutants have lower potency 
      and efficacy in chemotaxis and calcium mobilization assays using neutrophils. The 
      mutants also show lowered affinity to both IL-8 receptors A and B expressed 
      recombinantly in HL-60 cells and to neutrophils in [125I]IL-8 competition assays. 
      Additionally, the Leu25-->Tyr mutation introduces a novel monocyte 
      chemoattractant activity into IL-8. We therefore studied the displacement of 
      [125I]MIP-1 alpha by IL-8 Leu25-->Tyr from the CC-CKR-1 receptor. The mutant 
      displaces MIP-1 alpha ligand with an affinity only 12-fold less than MIP-1 alpha 
      itself. This suggests that mutations in this region of IL-8 are involved in 
      receptor binding and activation and in the control of specificity between CC and 
      CXC chemokines.
FAU - Lusti-Narasimhan, M
AU  - Lusti-Narasimhan M
AD  - Glaxo Institute for Molecular Biology, Plan-les-Ouates, Geneva, Switzerland.
FAU - Power, C A
AU  - Power CA
FAU - Allet, B
AU  - Allet B
FAU - Alouani, S
AU  - Alouani S
FAU - Bacon, K B
AU  - Bacon KB
FAU - Mermod, J J
AU  - Mermod JJ
FAU - Proudfoot, A E
AU  - Proudfoot AE
FAU - Wells, T N
AU  - Wells TN
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Chemokine CCL5)
RN  - 0 (Chemotactic Factors)
RN  - 0 (Cytokines)
RN  - 0 (DNA, Complementary)
RN  - 0 (Interleukin-8)
RN  - 0 (Lymphokines)
RN  - 0 (Monocyte Chemoattractant Proteins)
RN  - GMW67QNF9C (Leucine)
RN  - HG18B9YRS7 (Valine)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Amino Acid Sequence
MH  - Base Sequence
MH  - Calcium/metabolism
MH  - Chemokine CCL5
MH  - Chemotactic Factors/metabolism
MH  - Chemotaxis, Leukocyte/genetics
MH  - Cytokines/metabolism
MH  - DNA, Complementary
MH  - Humans
MH  - Interleukin-8/genetics/*metabolism
MH  - Leucine/genetics/*metabolism
MH  - Lymphokines/genetics/metabolism
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Monocyte Chemoattractant Proteins
MH  - Mutation
MH  - Valine/genetics/*metabolism
EDAT- 1995/02/10 00:00
MHDA- 1995/02/10 00:01
CRDT- 1995/02/10 00:00
PHST- 1995/02/10 00:00 [pubmed]
PHST- 1995/02/10 00:01 [medline]
PHST- 1995/02/10 00:00 [entrez]
AID - S0021-9258(17)30723-8 [pii]
AID - 10.1074/jbc.270.6.2716 [doi]
PST - ppublish
SO  - J Biol Chem. 1995 Feb 10;270(6):2716-21. doi: 10.1074/jbc.270.6.2716.