Rational design of tropoelastin peptide-based inhibitors of metalloproteinases.
Abnormal production of matrix metalloproteinases (MMPs) has been observed in a variety of diseases, such as emphysema, atherosclerosis, and cancer metastasis. Destruction of connective tissue ensues and elastin is often a key target. Three of the main elastolytic MMPs are the gelatinases MMP-2 and MMP-9 and the metalloelastase MMP-12. To investigate the possibility of using peptides to inhibit the elastolytic activity of these enzymes, we mapped the sites within tropoelastin recognized by MMP-9 and MMP-12. Peptides that correspond to regions overlapping these sites were then tested for their ability to inhibit these MMPs. These included an unmodified peptide directed against MMP-9 (peptide PP), cysteine-containing peptides that mimicked either the MMP-9 (peptide NCP) or the MMP-12 (peptide lin24) cleavage sites in tropoelastin and their cyclized forms (CP and cyc24, respectively), and a peptide containing a zinc-chelating hydroxamate group directed against MMP-9 (HP). The presence of a free sulfhydryl or hydroxamate group capable of chelating the zinc ion in the active site of the MMPs was generally found to increase the inhibitory activity of the peptides. The specificity of the inhibitors varied, with some of the inhibitors showing activity against all of the MMPs examined. None of the inhibitors had any significant effect on the activity of the unrelated serine protease, plasmin. K(i) values for the inhibitors were in the micromolar range. Our results suggest ways of developing other MMP inhibitors based on substrate recognition sites that may provide greater levels of inhibition.
Jensen
Sacha A
SA
School of Molecular and Microbial Biosciences G08, University of Sydney, NSW 2006, Sydney, Australia.
Andersen
Penny
P
Vrhovski
Bernadette
B
Weiss
Anthony S
AS
D000595:Amino Acid Sequence; D003545:Cysteine / Q000737:chemistry; D006801:Humans; D006877:Hydroxamic Acids / Q000737:chemistry; D053508:Matrix Metalloproteinase 12; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000737:chemistry; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D020033:Protein Isoforms / Q000138:chemical synthesis / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity; D014333:Tropoelastin / Q000138:chemical synthesis*
D006877:Hydroxamic Acids; D061965:Matrix Metalloproteinase Inhibitors; D010455:Peptides; D020033:Protein Isoforms; D014333:Tropoelastin; D020782:Matrix Metalloproteinases; D008666:Metalloendopeptidases; C507122:MMP12 protein, human; D053508:Matrix Metalloproteinase 12; D003545:Cysteine