PMID- 8077676
OWN - NLM
STAT- MEDLINE
DCOM- 19941005
LR  - 20131121
IS  - 0022-1767 (Print)
IS  - 0022-1767 (Linking)
VI  - 153
IP  - 6
DP  - 1994 Sep 15
TI  - The chemokines IL-8, monocyte chemoattractant protein-1, and I-309 are monomers 
      at physiologically relevant concentrations.
PG  - 2704-17
AB  - The chemokines are a family of immune mediators involved in a wide range of 
      inflammatory processes, most importantly as chemoattractants of monocytes, 
      neutrophils, lymphocytes, and fibroblasts to sites of inflammation. Nuclear 
      magnetic resonance and x-ray crystallographic studies have shown that IL-8 and 
      macrophage-inflammatory protein-1 beta (MIP-1 beta) form noncovalent dimers and 
      that platelet factor-4 (PF-4) forms noncovalent dimers and tetramers, leading to 
      the assumption that, as a family, the chemokines would form multimeric 
      structures. In this study, we analyze the association states of the chemokines 
      IL-8, monocyte chemoattractant protein-1 (MCP-1), and I-309, by using a 
      combination of size exclusion HPLC, sedimentation equilibrium 
      ultracentrifugation, and chemical cross-linking. We find that the association 
      states of MCP-1 and IL-8 are characterized by an equilibrium between monomers and 
      dimers: although dimers predominate at concentrations above 100 microM, these 
      chemokines are almost exclusively monomeric at the nanomolar concentrations at 
      which they display maximal chemotactic activity. I-309, by contrast, remains a 
      monomer at all concentrations tested. I-309 contains two additional cysteine 
      residues (C26 and C68) that are not found in any other members of the chemokine 
      family. We used cyanogen bromide and trypsin digestion strategies to demonstrate 
      that these two residues are linked in a unique intramolecular disulfide bond. 
      Furthermore, by using site-directed mutagenesis, we show that the integrity of 
      this bond is crucial for protein secretion.
FAU - Paolini, J F
AU  - Paolini JF
AD  - Department of Immunology, Duke University Medical Center, Durham, NC 27710.
FAU - Willard, D
AU  - Willard D
FAU - Consler, T
AU  - Consler T
FAU - Luther, M
AU  - Luther M
FAU - Krangel, M S
AU  - Krangel MS
LA  - eng
GR  - 5T32CA09058/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Immunol
JT  - Journal of immunology (Baltimore, Md. : 1950)
JID - 2985117R
RN  - 0 (Biopolymers)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Chemokines, CC)
RN  - 0 (Chemotactic Factors)
RN  - 0 (Cross-Linking Reagents)
RN  - 0 (Disulfides)
RN  - 0 (Interleukin-8)
RN  - EC 3.4.21.4 (Trypsin)
RN  - OS382OHJ8P (Cyanogen Bromide)
SB  - IM
EIN - J Immunol 1996 Apr 15;;156(8):following 3088
MH  - Amino Acid Sequence
MH  - Base Sequence
MH  - Biopolymers
MH  - Cell Line
MH  - Chemokine CCL2
MH  - *Chemokines, CC
MH  - Chemotactic Factors/*chemistry
MH  - Chromatography, Liquid
MH  - Cross-Linking Reagents
MH  - Cyanogen Bromide
MH  - Disulfides
MH  - Interleukin-8/*chemistry
MH  - Molecular Sequence Data
MH  - Protein Conformation
MH  - Transfection/methods
MH  - Trypsin
MH  - Ultracentrifugation
EDAT- 1994/09/15 00:00
MHDA- 1994/09/15 00:01
CRDT- 1994/09/15 00:00
PHST- 1994/09/15 00:00 [pubmed]
PHST- 1994/09/15 00:01 [medline]
PHST- 1994/09/15 00:00 [entrez]
PST - ppublish
SO  - J Immunol. 1994 Sep 15;153(6):2704-17.