PMID- 8573103
OWN - NLM
STAT- MEDLINE
DCOM- 19960301
LR  - 20190501
IS  - 0264-6021 (Print)
IS  - 1470-8728 (Electronic)
IS  - 0264-6021 (Linking)
VI  - 313 ( Pt 2)
IP  - Pt 2
DP  - 1996 Jan 15
TI  - Site-directed mutagenesis of monocyte chemoattractant protein-1 identifies two 
      regions of the polypeptide essential for biological activity.
PG  - 633-40
AB  - Monocyte chemoattractant protein-1 (MCP-1) mediates monocyte migration into 
      tissues in inflammatory diseases and atherosclerosis. We have investigated 
      structure-activity relationships for human MCP-1. Mutations were introduced based 
      upon differences between MCP-1 and the structurally related but functionally 
      distinct molecule interleukin-8 (IL-8). Mutant proteins produced using the 
      baculovirus/insect cell expression system were purified and their ability to 
      stimulate monocyte chemotaxis and elevation of intracellular calcium in THP-1 
      monocytic leukaemia cells was measured. Two regions in MCP-1 were identified as 
      important for its biological activity. One region consists of the sequence 
      Thr-Cys-Cys-Tyr (amino acids 10-13). Point mutations of Thr-10 to Arg and Tyr-13 
      to Ile greatly lowered MCP-1 activity. The second functionally important region 
      is formed by Ser-34 and Lys-35. Insertion of a Pro between these two residues, or 
      their substitution by the sequence Gly-Pro-His, caused nearly complete loss of 
      MCP-1 activity. Competition binding experiments showed that the mutations that 
      affected activity also lowered the ability to compete with wild-type MCP-1 for 
      receptors on THP-1 cells. Point mutations at positions 8, 15, 30, 37, 38 and 68 
      had little effect on MCP-1 activity. The important regions that we have 
      identified in MCP-1 correspond with previously identified functionally important 
      regions of IL-8, suggesting that the two molecules bind to their respective 
      receptors by similar contacts.
FAU - Beall, C J
AU  - Beall CJ
AD  - Neurobiotechnology Center, Ohio State University, Columbus 43210, USA.
FAU - Mahajan, S
AU  - Mahajan S
FAU - Kuhn, D E
AU  - Kuhn DE
FAU - Kolattukudy, P E
AU  - Kolattukudy PE
LA  - eng
GR  - HL48916/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (Chemokine CCL2)
RN  - 0 (DNA Primers)
RN  - 0 (Recombinant Proteins)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Baculoviridae/genetics
MH  - Base Sequence
MH  - Binding, Competitive
MH  - Cell Line
MH  - Chemokine CCL2/genetics/isolation & purification/*metabolism
MH  - Chromatography, Ion Exchange
MH  - DNA Primers
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Humans
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Protein Conformation
MH  - Recombinant Proteins/genetics/isolation & purification/metabolism
MH  - Spodoptera
PMC - PMC1216954
EDAT- 1996/01/15 00:00
MHDA- 1996/01/15 00:01
PMCR- 1996/07/15
CRDT- 1996/01/15 00:00
PHST- 1996/01/15 00:00 [pubmed]
PHST- 1996/01/15 00:01 [medline]
PHST- 1996/01/15 00:00 [entrez]
PHST- 1996/07/15 00:00 [pmc-release]
AID - 10.1042/bj3130633 [doi]
PST - ppublish
SO  - Biochem J. 1996 Jan 15;313 ( Pt 2)(Pt 2):633-40. doi: 10.1042/bj3130633.