title: De novo design of a single-chain diphenylporphyrin metalloprotein.
abstract: We describe the computational design of a single-chain four-helix bundle that noncovalently self-assembles with fully synthetic non-natural porphyrin cofactors. With this strategy, both the electronic structure of the cofactor as well as its protein environment may be varied to explore and modulate the functional and photophysical properties of the assembly. Solution characterization (NMR, UV-vis) of the protein showed that it bound with high specificity to the desired cofactors, suggesting that a uniquely structured protein and well-defined site had indeed been created. This provides a genetically expressed single-chain protein scaffold that will allow highly facile, flexible, and asymmetric variations to enable selective incorporation of different cofactors, surface-immobilization, and introduction of spectroscopic probes.
authors: Department of Biochemistry and Molecular Biophysics, Johnson Foundation, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
authors: Gretchen M
authors: GM
authors: Bender
authors: Andreas
authors: A
authors: Lehmann
authors: Hongling
authors: H
authors: Zou
authors: Hong
authors: H
authors: Cheng
authors: H Christopher
authors: HC
authors: Fry
authors: Don
authors: D
authors: Engel
authors: Michael J
authors: MJ
authors: Therien
authors: J Kent
authors: JK
authors: Blasie
authors: Heinrich
authors: H
authors: Roder
authors: Jeffrey G
authors: JG
authors: Saven
authors: William F
authors: WF
authors: DeGrado
mesh_terms: D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D002942:Circular Dichroism; D008667:Metalloproteins / Q000138:chemical synthesis / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D011166:Porphyrins / Q000138:chemical synthesis / Q000737:chemistry*; D017433:Protein Structure, Secondary; D013056:Spectrophotometry, Ultraviolet; D013816:Thermodynamics; D014461:Ultracentrifugation
chemical_list: D008667:Metalloproteins; D011166:Porphyrins
references: Biochemistry. 2000 Dec 5;39(48):14813-21
references: 11101297
references: J Phys Chem B. 2007 Feb 22;111(7):1823-33
references: 17256981
references: J Am Chem Soc. 2005 Apr 27;127(16):5804-5
references: 15839675
references: Proc Natl Acad Sci U S A. 2003 Nov 11;100(23):13140-5
references: 14595023
references: Annu Rev Biophys Biomol Struct. 2006;35:225-49
references: 16689635
references: Nano Lett. 2006 Nov;6(11):2387-94
references: 17090063
references: Curr Opin Chem Biol. 2003 Dec;7(6):741-8
references: 14644184
references: J Mol Biol. 2004 Apr 9;337(5):1195-205
references: 15046987
references: J Mol Biol. 2001 Feb 23;306(3):607-28
references: 11178917
references: J Mol Biol. 2003 Feb 28;326(4):1219-25
references: 12589764
references: J Mol Biol. 2001 Aug 31;311(5):1081-90
references: 11531341
references: Annu Rev Biochem. 2003;72:249-89
references: 12626339
references: Biochemistry. 1998 Mar 31;37(13):4635-43
references: 9521784
references: Nature. 1994 Mar 31;368(6470):425-32
references: 8133888
references: J Mol Biol. 2005 Mar 11;346(5):1441-54
references: 15713492
references: Curr Opin Struct Biol. 2001 Aug;11(4):485-90
references: 11495743
references: Chem Rev. 2001 Oct;101(10):3165-89
references: 11710067
references: J Phys Chem B. 1998 Apr 30;102(18):3586-616
references: 24889800
references: Protein Sci. 1994 Mar;3(3):522-4
references: 8019422
references: J Mol Biol. 2003 Dec 12;334(5):1101-15
references: 14643669
references: Annu Rev Biochem. 1999;68:779-819
references: 10872466
references: J Am Chem Soc. 2004 Jul 7;126(26):8141-7
references: 15225055
references: Chem Rev. 2004 Feb;104(2):617-49
references: 14871137
references: Anal Biochem. 1987 Feb 15;161(1):1-15
references: 3578775
references: Proc Natl Acad Sci U S A. 2004 Apr 13;101(15):5536-41
references: 15056758