title: A synthetic coiled-coil interactome provides heterospecific modules for molecular engineering.
abstract: The versatile coiled-coil protein motif is widely used to induce and control macromolecular interactions in biology and materials science. Yet the types of interaction patterns that can be constructed using known coiled coils are limited. Here we greatly expand the coiled-coil toolkit by measuring the complete pairwise interactions of 48 synthetic coiled coils and 7 human bZIP coiled coils using peptide microarrays. The resulting 55-member protein "interactome" includes 27 pairs of interacting peptides that preferentially heteroassociate. The 27 pairs can be used in combinations to assemble sets of 3 to 6 proteins that compose networks of varying topologies. Of special interest are heterospecific peptide pairs that participate in mutually orthogonal interactions. Such pairs provide the opportunity to dimerize two separate molecular systems without undesired crosstalk. Solution and structural characterization of two such sets of orthogonal heterodimers provide details of their interaction geometries. The orthogonal pair, along with the many other network motifs discovered in our screen, provide new capabilities for synthetic biology and other applications.
authors.affiliation: MIT, Department of Biology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Aaron W
authors.initials: AW
authors.lastname: Reinke
authors.forename: Robert A
authors.initials: RA
authors.lastname: Grant
authors.forename: Amy E
authors.initials: AE
authors.lastname: Keating
mesh_terms: D020816:Amino Acid Motifs; D050976:Basic-Leucine Zipper Transcription Factors / Q000138:chemical synthesis / Q000737:chemistry*; D006801:Humans; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D040081:Protein Array Analysis; D015202:Protein Engineering
chemical_list: D050976:Basic-Leucine Zipper Transcription Factors; D010455:Peptides
references.citation: Biochemistry. 2006 Sep 26;45(38):11324-32
references.pmid: 16981692
references.citation: Science. 1993 Nov 26;262(5138):1401-7
references.pmid: 8248779
references.citation: Proc Natl Acad Sci U S A. 2006 Jun 13;103(24):8989-94
references.pmid: 16754880
references.citation: Acta Crystallogr D Biol Crystallogr. 2002 Nov;58(Pt 11):1948-54
references.pmid: 12393927
references.citation: J Am Chem Soc. 2008 Sep 10;130(36):11921-7
references.pmid: 18710226
references.citation: Structure. 2002 Sep;10(9):1235-48
references.pmid: 12220495
references.citation: Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):3930-5
references.pmid: 10760265
references.citation: J Am Chem Soc. 2009 Jan 28;131(3):928-30
references.pmid: 19115943
references.citation: Nat Struct Biol. 2001 May;8(5):412-6
references.pmid: 11323715
references.citation: ACS Chem Biol. 2008 Jan 18;3(1):38-50
references.pmid: 18205291
references.citation: Proc Natl Acad Sci U S A. 2007 Jun 26;104(26):10853-8
references.pmid: 17567757
references.citation: Org Lett. 2008 Sep 4;10(17):3797-800
references.pmid: 18693746
references.citation: Biochemistry. 2003 Nov 25;42(46):13401-9
references.pmid: 14621985
references.citation: Nucleic Acids Res. 2007 Jul;35(Web Server issue):W375-83
references.pmid: 17452350
references.citation: Biochemistry. 2002 Apr 16;41(15):4866-71
references.pmid: 11939781
references.citation: Science. 1998 Jul 17;281(5375):389-92
references.pmid: 9665877
references.citation: Protein Sci. 2001 Mar;10(3):649-55
references.pmid: 11344333
references.citation: Biochemistry. 1967 Jul;6(7):1948-54
references.pmid: 6049437
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: Bioinformatics. 2006 May 1;22(9):1152-3
references.pmid: 16455747
references.citation: Methods Mol Biol. 2007;352:35-70
references.pmid: 17041258
references.citation: Nature. 2009 Apr 16;458(7240):859-64
references.pmid: 19370028
references.citation: Curr Opin Struct Biol. 2008 Aug;18(4):477-83
references.pmid: 18555680
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references.pmid: 15572765
references.citation: Curr Biol. 1993 Oct 1;3(10):658-67
references.pmid: 15335856
references.citation: Science. 2008 Mar 14;319(5869):1539-43
references.pmid: 18339942
references.citation: Mol Syst Biol. 2006;2:2006.2011
references.pmid: 16732192
references.citation: J Am Chem Soc. 2004 Sep 1;126(34):10514-5
references.pmid: 15327289
references.citation: Science. 2003 Jun 27;300(5628):2097-101
references.pmid: 12805554
references.citation: Cell. 1999 Oct 1;99(1):103-15
references.pmid: 10520998
references.citation: Proc Natl Acad Sci U S A. 2005 Sep 27;102(39):13891-6
references.pmid: 16172398
references.citation: Science. 2006 Mar 10;311(5766):1468-71
references.pmid: 16527982
references.citation: J Am Chem Soc. 2008 Jan 30;130(4):1321-7
references.pmid: 18171063