Engineered sarafotoxins as tissue inhibitor of metalloproteinases-like matrix metalloproteinase inhibitors.
The sarafotoxins and endothelins are approximately 25-residue peptides that spontaneously fold into a defined tertiary structure with specific pairing of four cysteines into two disulfide bonds. Their structures show an interesting topological similarity to the core of the metalloproteinase interaction sites of the tissue inhibitors of metalloproteinases. Previous work indicates that sarafotoxins and endothelins can be engineered to eliminate or greatly reduce their vasopressive action and that their structural framework can withstand multiple sequence changes. When sarafotoxin 6b, which possesses modest matrix metalloproteinase inhibitory activity, was C-terminally truncated to remove its toxic vasopressive activity, the metalloproteinase inhibitory activity was essentially abolished. However, further changes, based on the sequences of peptides selected from libraries of sarafotoxin variants or suggested by analogy with tissue inhibitors of metalloproteinases, progressively enhanced the matrix metalloproteinase inhibitory activity. Peptide variants with multiple substitutions folded correctly and formed native disulfide bonds. Improvements in matrix metalloproteinase affinity have generated a peptide with micromolar K(i) values for matrix metalloproteinase-1 and -9 that are selective inhibitors of different metalloproteinases. Characterization of its solution structure indicates a close similarity to sarafotoxin but with a more extended C-terminal helix. The effects of N-acetylation and other changes, as well as docking studies, support the hypothesis that the engineered sarafotoxins bind to matrix metalloproteinases in a manner analogous to the tissue inhibitors of metalloproteinases.
Lauer-Fields
Janelle L
JL
College of Biomedical Science, Florida Atlantic University, Boca Raton, Florida 33431; Department of Chemistry & Biochemistry, Florida Atlantic University, Boca Raton, Florida 33431.
Cudic
Mare
M
Department of Chemistry & Biochemistry, Florida Atlantic University, Boca Raton, Florida 33431.
Wei
Shuo
S
College of Biomedical Science, Florida Atlantic University, Boca Raton, Florida 33431.
Mari
Frank
F
Department of Chemistry & Biochemistry, Florida Atlantic University, Boca Raton, Florida 33431.
Fields
Gregg B
GB
Department of Chemistry & Biochemistry, Florida Atlantic University, Boca Raton, Florida 33431.
Brew
Keith
K
College of Biomedical Science, Florida Atlantic University, Boca Raton, Florida 33431. Electronic address: kbrew@fau.edu.
D000595:Amino Acid Sequence; D003461:Crystallography; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D015202:Protein Engineering; D013329:Structure-Activity Relationship; D019714:Tissue Inhibitor of Metalloproteinases / Q000737:chemistry / Q000494:pharmacology*; D014757:Viper Venoms / Q000737:chemistry / Q000494:pharmacology*
D019714:Tissue Inhibitor of Metalloproteinases; D014757:Viper Venoms; C057354:sarafotoxins s6