De novo design of a single-chain diphenylporphyrin metalloprotein.
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.
Bender
Gretchen M
GM
Department of Biochemistry and Molecular Biophysics, Johnson Foundation, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Lehmann
Andreas
A
Zou
Hongling
H
Cheng
Hong
H
Fry
H Christopher
HC
Engel
Don
D
Therien
Michael J
MJ
Blasie
J Kent
JK
Roder
Heinrich
H
Saven
Jeffrey G
JG
DeGrado
William F
WF
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
D008667:Metalloproteins; D011166:Porphyrins
Biochemistry. 2000 Dec 5;39(48):14813-21
11101297
J Phys Chem B. 2007 Feb 22;111(7):1823-33
17256981
J Am Chem Soc. 2005 Apr 27;127(16):5804-5
15839675
Proc Natl Acad Sci U S A. 2003 Nov 11;100(23):13140-5
14595023
Annu Rev Biophys Biomol Struct. 2006;35:225-49
16689635
Nano Lett. 2006 Nov;6(11):2387-94
17090063
Curr Opin Chem Biol. 2003 Dec;7(6):741-8
14644184
J Mol Biol. 2004 Apr 9;337(5):1195-205
15046987
J Mol Biol. 2001 Feb 23;306(3):607-28
11178917
J Mol Biol. 2003 Feb 28;326(4):1219-25
12589764
J Mol Biol. 2001 Aug 31;311(5):1081-90
11531341
Annu Rev Biochem. 2003;72:249-89
12626339
Biochemistry. 1998 Mar 31;37(13):4635-43
9521784
Nature. 1994 Mar 31;368(6470):425-32
8133888
J Mol Biol. 2005 Mar 11;346(5):1441-54
15713492
Curr Opin Struct Biol. 2001 Aug;11(4):485-90
11495743
Chem Rev. 2001 Oct;101(10):3165-89
11710067
J Phys Chem B. 1998 Apr 30;102(18):3586-616
24889800
Protein Sci. 1994 Mar;3(3):522-4
8019422
J Mol Biol. 2003 Dec 12;334(5):1101-15
14643669
Annu Rev Biochem. 1999;68:779-819
10872466
J Am Chem Soc. 2004 Jul 7;126(26):8141-7
15225055
Chem Rev. 2004 Feb;104(2):617-49
14871137
Anal Biochem. 1987 Feb 15;161(1):1-15
3578775
Proc Natl Acad Sci U S A. 2004 Apr 13;101(15):5536-41
15056758