Conversion of monocyte chemoattractant protein-1 into a neutrophil attractant by substitution of two amino acids.
The small cytokine monocyte chemoattractant protein-1 has structural similarity to the neutrophil chemoattractant interleukin-8, but each protein is specific in attracting its own target cell. To investigate the structural basis of this cell type specificity, we have developed an Escherichia coli expression system for the monocyte chemoattractant and mutagenized selected amino acid residues to ones found at the corresponding positions of interleukin-8. We find that a double mutation of tyrosine 28 and arginine 30 to leucine and valine, respectively, causes a drastic decrease in chemotactic activity toward monocytes with the appearance of a novel (interleukin-8-like) neutrophil chemotactic activity. Computer graphic analysis predicts that, with the double substitution, a putative receptor binding groove of the monocyte chemoattractant protein would become topographically similar to that of interleukin-8. We therefore postulate that one or both of these amino acid residues are part of the binding contact of these small cytokines and their receptors.
Beall
C J
CJ
Biotechnology Center, Ohio State University, Columbus 43210.
Mahajan
S
S
Kolattukudy
P E
PE
D000595:Amino Acid Sequence; D001483:Base Sequence; D018932:Chemokine CCL2; D002630:Chemotactic Factors / Q000737:chemistry / Q000235:genetics*; D002634:Chemotaxis, Leukocyte; D003001:Cloning, Molecular; D003198:Computer Simulation; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004730:Endothelium, Vascular / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D006801:Humans; D016209:Interleukin-8 / Q000737:chemistry / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009000:Monocytes / Q000502:physiology; D009504:Neutrophils / Q000502:physiology; D009838:Oligodeoxyribonucleotides; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry
D018932:Chemokine CCL2; D002630:Chemotactic Factors; D016209:Interleukin-8; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D004247:DNA