title: Design of a bioactive cell-penetrating peptide: when a transduction domain does more than transduce.
abstract: The discovery of cell-penetrating peptides (CPPs) has facilitated delivery of peptides into cells to affect cellular behavior. Previously, we were successful at developing a phosphopeptide mimetic of the small heat shock-like protein HSP20 . Building on this success we developed a cell-permeant peptide inhibitor of mitogen-activated protein kinase-activated protein kinase 2 (MK2). It is well documented that inhibition of MK2 may be beneficial for a myriad of human diseases including those involving inflammation and fibrosis. During the optimization of the activity and specificity of the MK2 inhibitor (MK2i) we closely examined the effect of cell-penetrating peptide identity. Surprisingly, the identity of the CPP dictated kinase specificity and functional activity to an extent that rivaled that of the therapeutic peptide. The results reported herein have wide implications for delivering therapeutics with CPPs and indicate that judicious choice of CPP is crucial to the ultimate therapeutic success.
authors.affiliation: Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN 47907-2032, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Ward
authors.forename: Brandon L
authors.initials: BL
authors.lastname: Seal
authors.forename: Colleen M
authors.initials: CM
authors.lastname: Brophy
authors.forename: Alyssa
authors.initials: A
authors.lastname: Panitch
mesh_terms: D000595:Amino Acid Sequence; D002470:Cell Survival; D002478:Cells, Cultured; D015195:Drug Design; D004847:Epithelial Cells; D050886:HSP20 Heat-Shock Proteins / Q000737:chemistry*; D006801:Humans; D020128:Inhibitory Concentration 50; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors*; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011494:Protein Kinases / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors*; D000099190:MAP-Kinase-Activated Kinase 2
chemical_list: D050886:HSP20 Heat-Shock Proteins; D047908:Intracellular Signaling Peptides and Proteins; D010455:Peptides; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases; D000099190:MAP-Kinase-Activated Kinase 2
references.citation: J Biol Chem. 2001 Feb 23;276(8):5836-40
references.pmid: 11084031
references.citation: Blood. 1994 Apr 1;83(7):1791-8
references.pmid: 8142647
references.citation: Cell Signal. 2000 Jan;12(1):1-13
references.pmid: 10676842
references.citation: J Biol Chem. 1994 Apr 8;269(14):10444-50
references.pmid: 8144628
references.citation: Gene Ther. 2002 Nov;9(21):1422-8
references.pmid: 12378404
references.citation: FEBS Lett. 1996 Oct 21;395(2-3):133-6
references.pmid: 8898080
references.citation: Cell. 1994 Sep 23;78(6):1027-37
references.pmid: 7923353
references.citation: Biochem J. 1993 Dec 15;296 ( Pt 3):843-9
references.pmid: 8280084
references.citation: Curr Med Chem. 2007;14(21):2214-34
references.pmid: 17896971
references.citation: Bioconjug Chem. 2004 May-Jun;15(3):482-90
references.pmid: 15149175
references.citation: Nat Biotechnol. 2005 Mar;23(3):329-36
references.pmid: 15711537
references.citation: Biochem Biophys Res Commun. 2009 May 8;382(3):535-9
references.pmid: 19289101
references.citation: J Exp Med. 2007 Jul 9;204(7):1637-52
references.pmid: 17576778
references.citation: Biochem Soc Trans. 2002 Nov;30(Pt 6):959-63
references.pmid: 12440954
references.citation: FEBS Lett. 1996 Sep 2;392(3):209-14
references.pmid: 8774846
references.citation: Eur J Neurosci. 2008 Aug;28(4):642-54
references.pmid: 18702688
references.citation: Leukemia. 1995 Feb;9(2):288-94
references.pmid: 7869766
references.citation: Biochem J. 2003 Apr 1;371(Pt 1):199-204
references.pmid: 12534346
references.citation: Methods. 2001 Jul;24(3):247-56
references.pmid: 11403574
references.citation: Cell. 1988 Dec 23;55(6):1189-93
references.pmid: 2849510
references.citation: J Biol Chem. 2002 Nov 8;277(45):42596-602
references.pmid: 12202485
references.citation: J Exp Med. 2002 Dec 16;196(12):1563-74
references.pmid: 12486099
references.citation: Nat Methods. 2007 Feb;4(2):119-20
references.pmid: 17264857
references.citation: J Biol Chem. 1995 Jun 16;270(24):14255-8
references.pmid: 7782278
references.citation: J Pept Res. 2000 Nov;56(5):318-25
references.pmid: 11095185
references.citation: J Immunol. 2006 Aug 1;177(3):1913-7
references.pmid: 16849504
references.citation: Trends Mol Med. 2007 Oct;13(10):443-8
references.pmid: 17913584
references.citation: Front Biosci. 2008 May 01;13:5511-21
references.pmid: 18508601
references.citation: J Cell Sci. 1997 Feb;110 ( Pt 3):357-68
references.pmid: 9057088
references.citation: EMBO J. 2002 Oct 15;21(20):5396-407
references.pmid: 12374740
references.citation: Arch Biochem Biophys. 1996 Oct 15;334(2):206-13
references.pmid: 8900394
references.citation: Bioconjug Chem. 2004 Nov-Dec;15(6):1246-53
references.pmid: 15546190
references.citation: Mol Cell. 2005 Jan 7;17(1):37-48
references.pmid: 15629715
references.citation: Nat Rev Mol Cell Biol. 2006 Feb;7(2):120-30
references.pmid: 16421520
references.citation: Biochemistry. 2002 Oct 22;41(42):12652-61
references.pmid: 12379107
references.citation: J Med Chem. 2002 Aug 15;45(17):3612-8
references.pmid: 12166934
references.citation: Biochem Biophys Res Commun. 1994 Apr 29;200(2):1118-24
references.pmid: 8179591
references.citation: J Immunol. 2002 May 1;168(9):4667-73
references.pmid: 11971016
references.citation: J Biol Chem. 1996 Apr 5;271(14):8488-92
references.pmid: 8626550
references.citation: Am J Physiol Gastrointest Liver Physiol. 2006 Jun;290(6):G1298-306
references.pmid: 16423921
references.citation: FASEB J. 2003 Jul;17(10):1358-60
references.pmid: 12738803
references.citation: Nat Cell Biol. 2004 Mar;6(3):189-96
references.pmid: 15039791
references.citation: FASEB J. 2005 Feb;19(2):261-3
references.pmid: 15598710
references.citation: Biochem Pharmacol. 1997 May 9;53(9):1239-47
references.pmid: 9214684
references.citation: Trends Cell Biol. 2000 Jul;10(7):290-5
references.pmid: 10856932
references.citation: Proc Natl Acad Sci U S A. 2000 Dec 19;97(26):14772-7
references.pmid: 11121077
references.citation: Nat Biotechnol. 2008 Jan;26(1):127-32
references.pmid: 18183025
references.citation: J Med Chem. 1999 Mar 11;42(5):910-9
references.pmid: 10072688
references.citation: Biomacromolecules. 2003 Nov-Dec;4(6):1572-82
references.pmid: 14606882
references.citation: J Biol Chem. 2006 Aug 18;281(33):23658-67
references.pmid: 16774924
references.citation: Cancer Res. 2001 Jan 15;61(2):474-7
references.pmid: 11212234
references.citation: Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1864-8
references.pmid: 1672046
references.citation: Biochem J. 2000 Oct 1;351(Pt 1):95-105
references.pmid: 10998351
references.citation: Proc Natl Acad Sci U S A. 2000 Nov 21;97(24):13003-8
references.pmid: 11087855
references.citation: Cancer Cell. 2007 Feb;11(2):175-89
references.pmid: 17292828
references.citation: Cell. 1988 Dec 23;55(6):1179-88
references.pmid: 2849509
references.citation: Curr Opin Biotechnol. 2002 Feb;13(1):52-6
references.pmid: 11849958
references.citation: J Biol Chem. 2002 Oct 4;277(40):37401-5
references.pmid: 12171911
references.citation: Nat Cell Biol. 1999 Jun;1(2):94-7
references.pmid: 10559880
references.citation: Mol Pharmacol. 1994 Dec;46(6):1077-83
references.pmid: 7808427
references.citation: Circ Res. 2007 Nov 26;101(11):1104-12
references.pmid: 17885219
references.citation: Microbiol Mol Biol Rev. 2004 Jun;68(2):320-44
references.pmid: 15187187
references.citation: Curr Pharm Des. 2005;11(28):3597-611
references.pmid: 16305497
references.citation: Trends Cell Biol. 1998 Feb;8(2):84-7
references.pmid: 9695814
references.citation: Arch Biochem Biophys. 2002 Jan 15;397(2):224-31
references.pmid: 11795875
references.citation: J Cell Biochem. 1995 Feb;57(2):321-30
references.pmid: 7759569