title: Genetic incorporation of a small, environmentally sensitive, fluorescent probe into proteins in Saccharomyces cerevisiae.
abstract: Here, we report that the fluorescent amino acid, 3-(6-acetylnaphthalen-2-ylamino)-2-aminopropanoic acid (Anap), can be genetically incorporated into proteins in yeast with excellent selectivity and efficiency by means of an orthogonal tRNA/aminoacyl-tRNA synthetase pair. This small, environmentally sensitive fluorophore was site-specifically incorporated into Escherichia coli glutamine binding protein and used to directly probe local structural changes caused by ligand binding. The small size of Anap and the ability to introduce it by simple mutagenesis at defined sites in the proteome make it a useful local probe of protein structure, molecular interactions, protein folding, and localization.
authors.affiliation: Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
authors.forename: Hyun Soo
authors.initials: HS
authors.lastname: Lee
authors.forename: Jiantao
authors.initials: J
authors.lastname: Guo
authors.forename: Edward A
authors.initials: EA
authors.lastname: Lemke
authors.forename: Romerson D
authors.initials: RD
authors.lastname: Dimla
authors.forename: Peter G
authors.initials: PG
authors.lastname: Schultz
mesh_terms: D026921:Amino Acid Transport Systems, Neutral / Q000235:genetics; D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics; D005456:Fluorescent Dyes / Q000737:chemistry*; D015202:Protein Engineering; D012343:RNA, Transfer / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D015091:beta-Alanine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics
chemical_list: C546053:3-(6-acetylnaphthalen-2-ylamino)-2-aminopropanoic acid; D026921:Amino Acid Transport Systems, Neutral; D029968:Escherichia coli Proteins; D005456:Fluorescent Dyes; D029701:Saccharomyces cerevisiae Proteins; C528486:glnH protein, E coli; D015091:beta-Alanine; D012343:RNA, Transfer; D000604:Amino Acyl-tRNA Synthetases
references.citation: Science. 2000 Sep 8;289(5485):1760-3
references.pmid: 10976071
references.citation: Methods Enzymol. 2000;327:565-78
references.pmid: 11045009
references.citation: Nat Chem Biol. 2007 Jun;3(6):321-2
references.pmid: 17450134
references.citation: Science. 2002 Apr 19;296(5567):503-7
references.pmid: 11964472
references.citation: Biochemistry. 1979 Jul 10;18(14):3075-8
references.pmid: 465454
references.citation: Science. 2006 Apr 14;312(5771):217-24
references.pmid: 16614209
references.citation: Science. 2003 Aug 15;301(5635):964-7
references.pmid: 12920298
references.citation: Chem Biol. 2000 Apr;7(4):253-61
references.pmid: 10780925
references.citation: Science. 2002 May 31;296(5573):1700-3
references.pmid: 12040199
references.citation: Nat Biotechnol. 1999 Oct;17(10):969-73
references.pmid: 10504696
references.citation: J Am Chem Soc. 2006 Jul 12;128(27):8738-9
references.pmid: 16819861
references.citation: Chem Biol. 2003 Jun;10(6):511-9
references.pmid: 12837384
references.citation: Proc Natl Acad Sci U S A. 2006 Jun 27;103(26):9785-9
references.pmid: 16785423
references.citation: J Mol Biol. 1998 Apr 24;278(1):219-29
references.pmid: 9571045
references.citation: Nat Biotechnol. 2003 Jan;21(1):86-9
references.pmid: 12469133
references.citation: J Phys Chem A. 2005 Feb 24;109(7):1295-8
references.pmid: 16833443
references.citation: J Am Chem Soc. 2002 May 29;124(21):6063-76
references.pmid: 12022841
references.citation: Nat Methods. 2005 Feb;2(2):99-104
references.pmid: 15782206
references.citation: Nat Rev Mol Cell Biol. 2002 Dec;3(12):906-18
references.pmid: 12461557
references.citation: Nature. 1986 Jan 2-8;319(6048):70-3
references.pmid: 3941741
references.citation: Biochemistry. 1986 Jun 3;25(11):3479-86
references.pmid: 3755359
references.citation: J Am Chem Soc. 2003 Dec 31;125(52):16228-34
references.pmid: 14692764
references.citation: Nat Biotechnol. 2003 May;21(5):539-45
references.pmid: 12692560
references.citation: Proc Natl Acad Sci U S A. 2001 Dec 18;98(26):15003-8
references.pmid: 11752449
references.citation: Bioorg Med Chem. 2005 May 16;13(10):3379-84
references.pmid: 15848750
references.citation: Nature. 1997 Aug 28;388(6645):882-7
references.pmid: 9278050
references.citation: Nat Methods. 2007 Mar;4(3):239-44
references.pmid: 17322890
references.citation: Angew Chem Int Ed Engl. 2007;46(48):9239-42
references.pmid: 17893898
references.citation: J Am Chem Soc. 2004 Nov 10;126(44):14306-7
references.pmid: 15521721
references.citation: J Am Chem Soc. 2008 May 14;130(19):6066-7
references.pmid: 18426210
references.citation: EMBO J. 2000 May 15;19(10):2351-61
references.pmid: 10811626
references.citation: Science. 1998 Jul 10;281(5374):269-72
references.pmid: 9657724