Structure--activity relationships in human interleukin-1{alpha}: identification of key residues for expression of biological activities

To identify the sites important for the different biologicalactivities of human interleukin-l (hIL-1), 56 single-amino acid-substitutedmutants of hIL-l were produced in Escherichia coli using site-directedmutagenesis, and were examined for their biological activitiessuch as mouse lymphocyte activating factor activity (LAF activity),cytostatic activity against human melanoma cells A-375 (A375activity) and prostaglandin E2 (PGE2) inducing activity in humanosteosarcoma cells MG-63 (PEI activity). Two amino acid residues,Asp26 and Asp151, were found to be important for these activities.The replacement of Asp26 by Val caused a decrease in LAF andA375 activities by one or two orders of magnitude and a slightdecrease in A375 activity. The Tyr or Phe substitution for Asp151caused decreases in LAF and A375 activities by one or two ordersof magnitude and complete loss of PEI activity. The change fromAsp151 to Lys or Arg resulted in marked decrease in LAF activityand complete loss of A375 and PEI activities. Since Asp26 andAsp151 are close to each other in the three-dimensional structure,the region involving these amino acids seems to be importantfor the biological activities of hIL-1.