title: Insertion of an elastase-binding loop into interleukin-1 beta.
abstract: The protease-binding sequence EAIPMSIPPE from alpha 1-antitrypsin has been inserted into the cytokine interleukin-1 beta, replacing residues 50-53. The resulting mutant protein was cleaved specifically at a single site by elastase and chymotrypsin, but not by trypsin. The cleavage by elastase was shown to be between Met and Ser of the inserted loop. In contrast, wild-type interleukin is not susceptible to cleavage by any of these enzymes. The mutant protein acts as an inhibitor of elastase, with a KI of approximately 30 microM. The wild type displays no such inhibitory activity. The overall structure of the mutant, as demonstrated by CD, appears to be indistinguishable from that of the wild type. These results indicate that the protease-binding region of alpha 1-antitrypsin can be recognized and is active even within the context of an entirely different protein structure. Given that interleukin-1 beta binds to, and is internalized by, many types of cells, this hybrid protein also demonstrates the feasibility of using interleukin-1 beta as a delivery system for useful therapeutic agents.
authors.affiliation: Brandeis University, Rosentiel Basic Medical Research Center, Waltham, MA 02254.
authors.forename: A J
authors.initials: AJ
authors.lastname: Wolfson
authors.forename: M
authors.initials: M
authors.lastname: Kanaoka
authors.forename: F T
authors.initials: FT
authors.lastname: Lau
authors.forename: D
authors.initials: D
authors.lastname: Ringe
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D006801:Humans; D007375:Interleukin-1 / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D015345:Oligonucleotide Probes; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors / Q000235:genetics*; D011480:Protease Inhibitors / Q000737:chemistry; D011487:Protein Conformation; D000515:alpha 1-Antitrypsin / Q000235:genetics
chemical_list: D007375:Interleukin-1; D015345:Oligonucleotide Probes; D011480:Protease Inhibitors; D000515:alpha 1-Antitrypsin; D010196:Pancreatic Elastase