PMID- 9688530
OWN - NLM
STAT- MEDLINE
DCOM- 19980813
LR  - 20190621
IS  - 0014-5793 (Print)
IS  - 0014-5793 (Linking)
VI  - 430
IP  - 3
DP  - 1998 Jul 3
TI  - Mapping of MCP-1 functional domains by peptide analysis and site-directed 
      mutagenesis.
PG  - 158-64
AB  - Monocyte chemoattractant protein-1 (MCP-1) is a member of the beta chemokine 
      family which acts through specific seven transmembrane receptors to recruit 
      monocytes, basophils, and T lymphocytes to sites of inflammation. To identify 
      regions of the human MCP-1 protein which are important for its biological 
      activity, we have synthesized domain-specific peptides and tested their ability 
      to antagonize MCP-1 binding and chemotaxis in THP-1 cells. We have found that an 
      intercysteine first loop peptide encompassing amino acids 13-35 inhibits MCP-1 
      binding and chemotactic activity, while peptides representing the amino-terminus 
      (amino acids 1-10), second loop (amino acids 37-51), and carboxy-terminus (amino 
      acids 56-71) of MCP-1 have no effect. In addition, we have found that cyclization 
      of the first loop peptide by disulfide linkage and blocking the C-terminus of the 
      peptide by amidation increases the activity of this peptide to block MCP-1 
      binding and chemotaxis. In order to specifically identify amino acid residues 
      within the first loop that are crucial for MCP-1 functional activity, we have 
      substituted alanine for tyrosine (Y13A) or arginine (R18A) in MCP-1 recombinant 
      proteins. While baculovirus produced wild type and R18A MCP-1 proteins are 
      indistinguishable in their ability to induce THP-1 chemotaxis and show modest 
      effects in binding activity compared to commercially available recombinant MCP-1 
      protein, the Y13A point mutation causes a dramatic loss in function. The 
      identification of functional domains of MCP-1 will assist in the design of MCP-1 
      receptor antagonists which may be clinically beneficial in a number of 
      inflammatory diseases.
FAU - Steitz, S A
AU  - Steitz SA
AD  - Tanabe Research Laboratories USA, Inc., San Diego, CA 92121, USA.
FAU - Hasegawa, K
AU  - Hasegawa K
FAU - Chiang, S L
AU  - Chiang SL
FAU - Cobb, R R
AU  - Cobb RR
FAU - Castro, M A
AU  - Castro MA
FAU - Lobl, T J
AU  - Lobl TJ
FAU - Yamada, M
AU  - Yamada M
FAU - Lazarides, E
AU  - Lazarides E
FAU - Cardarelli, P M
AU  - Cardarelli PM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - FEBS Lett
JT  - FEBS letters
JID - 0155157
RN  - 0 (Chemokine CCL2)
RN  - 0 (Peptides)
RN  - 0 (Peptides, Cyclic)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 42HK56048U (Tyrosine)
RN  - 94ZLA3W45F (Arginine)
SB  - IM
MH  - Amino Acid Sequence
MH  - Arginine/physiology
MH  - Binding, Competitive
MH  - Cell Line
MH  - Chemokine CCL2/*chemistry/genetics
MH  - Chemotaxis, Leukocyte/*physiology
MH  - Humans
MH  - Molecular Sequence Data
MH  - Monocytes/cytology/metabolism
MH  - Mutagenesis, Site-Directed
MH  - Mutation
MH  - Peptides/chemical synthesis/*pharmacology
MH  - Peptides, Cyclic/chemical synthesis/pharmacology
MH  - Recombinant Fusion Proteins
MH  - Tyrosine/physiology
EDAT- 1998/08/04 00:00
MHDA- 1998/08/04 00:01
CRDT- 1998/08/04 00:00
PHST- 1998/08/04 00:00 [pubmed]
PHST- 1998/08/04 00:01 [medline]
PHST- 1998/08/04 00:00 [entrez]
AID - S0014-5793(98)00637-1 [pii]
AID - 10.1016/s0014-5793(98)00637-1 [doi]
PST - ppublish
SO  - FEBS Lett. 1998 Jul 3;430(3):158-64. doi: 10.1016/s0014-5793(98)00637-1.