PMID- 10587439
OWN - NLM
STAT- MEDLINE
DCOM- 20000110
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 38
IP  - 49
DP  - 1999 Dec 7
TI  - Identification of surface residues of the monocyte chemotactic protein 1 that
      affect signaling through the receptor CCR2.
PG  - 16167-77
AB  - The CC chemokine, monocyte chemotactic protein, 1 (MCP-1) functions as a major
      chemoattractant for T-cells and monocytes by interacting with the
      seven-transmembrane G protein-coupled receptor CCR2. To identify which residues
      of MCP-1 contribute to signaling though CCR2, we mutated all the surface-exposed 
      residues to alanine and other amino acids and made some selective large changes
      at the amino terminus. We then characterized the impact of these mutations on
      three postreceptor pathways involving inhibition of cAMP synthesis, stimulation
      of cytosolic calcium influx, and chemotaxis. The results highlight several
      important features of the signaling process and the correlation between binding
      and signaling: The amino terminus of MCP-1 is essential as truncation of residues
      2-8 ([1+9-76]hMCP-1) results in a protein that cannot stimulate chemotaxis.
      However, the exact peptide sequence may be unimportant as individual alanine
      mutations or simultaneous replacement of residues 3-6 with alanine had little
      effect. Y13 is also important and must be a large nonpolar residue for chemotaxis
      to occur. Interestingly, both Y13 and [1+9-76]hMCP-1 are high-affinity binders
      and thus affinity of these mutants is not correlated with ability to promote
      chemotaxis. For the other surface residues there is a strong correlation between 
      binding affinity and agonist potency in all three signaling pathways. Perhaps the
      most interesting observation is that although Y13A and [1+9-76]hMCP are
      antagonists of chemotaxis, they are agonists of pathways involving inhibition of 
      cAMP synthesis and, in the case of Y13A, calcium influx. These results
      demonstrate that these two well-known signaling events are not sufficient to
      drive chemotaxis. Furthermore, it suggests that specific molecular features of
      MCP-1 induce different conformations in CCR2 that are coupled to separate
      postreceptor pathways. Therefore, by judicious design of antagonists, it should
      be possible to trap CCR2 in conformational states that are unable to stimulate
      all of the pathways required for chemotaxis.
FAU - Jarnagin, K
AU  - Jarnagin K
AD  - Department of Molecular and Cell Biology, University of California at Berkeley
      94720, USA. kjarnagin@iconixpharm.com
FAU - Grunberger, D
AU  - Grunberger D
FAU - Mulkins, M
AU  - Mulkins M
FAU - Wong, B
AU  - Wong B
FAU - Hemmerich, S
AU  - Hemmerich S
FAU - Paavola, C
AU  - Paavola C
FAU - Bloom, A
AU  - Bloom A
FAU - Bhakta, S
AU  - Bhakta S
FAU - Diehl, F
AU  - Diehl F
FAU - Freedman, R
AU  - Freedman R
FAU - McCarley, D
AU  - McCarley D
FAU - Polsky, I
AU  - Polsky I
FAU - Ping-Tsou, A
AU  - Ping-Tsou A
FAU - Kosaka, A
AU  - Kosaka A
FAU - Handel, T M
AU  - Handel TM
LA  - eng
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  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Amino Acids)
RN  - 0 (CCR2 protein, human)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Peptide Fragments)
RN  - 0 (Receptors, CCR2)
RN  - 0 (Receptors, Chemokine)
RN  - 0 (Receptors, Cytokine)
RN  - 42HK56048U (Tyrosine)
RN  - E0399OZS9N (Cyclic AMP)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Amino Acids/isolation & purification/*physiology
MH  - Binding Sites/genetics
MH  - Calcium/antagonists & inhibitors/metabolism
MH  - Cell Line
MH  - Cell Membrane/genetics/physiology
MH  - Cell Migration Inhibition
MH  - Chemokine CCL2/agonists/genetics/*physiology
MH  - Cyclic AMP/antagonists & inhibitors
MH  - Humans
MH  - Peptide Fragments/genetics/metabolism/physiology
MH  - Protein Structure, Secondary/genetics
MH  - Protein Structure, Tertiary/genetics
MH  - Receptors, CCR2
MH  - Receptors, Chemokine/metabolism/*physiology
MH  - Receptors, Cytokine/metabolism/*physiology
MH  - *Signal Transduction/genetics
MH  - Tyrosine/genetics/physiology
EDAT- 1999/12/10 00:00
MHDA- 1999/12/10 00:01
CRDT- 1999/12/10 00:00
PHST- 1999/12/10 00:00 [pubmed]
PHST- 1999/12/10 00:01 [medline]
PHST- 1999/12/10 00:00 [entrez]
AID - bi9912239 [pii]
AID - 10.1021/bi9912239 [doi]
PST - ppublish
SO  - Biochemistry. 1999 Dec 7;38(49):16167-77. doi: 10.1021/bi9912239.