title: Inhibition of furin by polyarginine-containing peptides: nanomolar inhibition by nona-D-arginine.
abstract: Polyarginine-containing peptides represent potent inhibitors of furin, a mammalian endoprotease that plays an important role in metabolism, activation of pathogenic toxins, and viral proliferation. The therapeutic use of D-polyarginines is especially interesting because they are not cleaved by furin and possess inhibitory potency almost equal to L-polyarginines. In this study we attempted to determine the important elements within polyarginines that contribute to effective inhibition. Structure-function analyses of polyarginine peptides showed that inhibition by polyarginine-containing peptides appeared to depend on the total number of basic charges of the positively charged inhibitors bound to the negatively charged substrate binding pocket; peptide positioning did not appear to be rigorously determined. Screening of L- and D-decapeptide positional scanning combinatorial peptide libraries indicated a preference for basic residues in nearly all positions, similar to previous results with hexapeptide libraries. Length and terminal modification studies showed that the most potent D-polyarginine tested was nona-D-arginine (D9R) amide with a K(i) of 1.3 nm. D9R amide was shown to protect RAW264.7 cells against anthrax toxemia with an IC(50) of 3.7 microm. Because of its high stability, specificity, low toxicity, small molecular weight, and extremely low K(i) against furin, D9R amide or its derivatives may represent promising compounds for therapeutic use.
authors.affiliation: Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center, 1901 Perdido Street, New Orleans, LA 70112, USA.
authors.forename: Magdalena M
authors.initials: MM
authors.lastname: Kacprzak
authors.forename: Juan R
authors.initials: JR
authors.lastname: Peinado
authors.forename: Manuel E
authors.initials: ME
authors.lastname: Than
authors.forename: Jon
authors.initials: J
authors.lastname: Appel
authors.forename: Stefan
authors.initials: S
authors.lastname: Henrich
authors.forename: Gregory
authors.initials: G
authors.lastname: Lipkind
authors.forename: Richard A
authors.initials: RA
authors.lastname: Houghten
authors.forename: Wolfram
authors.initials: W
authors.lastname: Bode
authors.forename: Iris
authors.initials: I
authors.lastname: Lindberg
mesh_terms: D000409:Alanine / Q000737:chemistry; D000818:Animals; D000942:Antigens, Bacterial / Q000378:metabolism; D001120:Arginine / Q000737:chemistry*; D001427:Bacterial Toxins / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D018360:Crystallography, X-Ray; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D045683:Furin / Q000037:antagonists & inhibitors*; D006801:Humans; D006868:Hydrolysis; D020128:Inhibitory Concentration 50; D007477:Ions; D007700:Kinetics; D008239:Lysine / Q000737:chemistry; D051379:Mice; D008958:Models, Molecular; D009842:Oligopeptides / Q000494:pharmacology; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D018076:DNA, Complementary; D007477:Ions; D009842:Oligopeptides; D019151:Peptide Library; D010455:Peptides; C030325:anthrax toxin; C491427:nonaarginine; C015462:polyarginine; D001120:Arginine; D045683:Furin; D008239:Lysine; D000409:Alanine