Expressed protein ligation: a resourceful tool to study protein  structure and function.
This review outlines the use of expressed protein ligation (EPL) to study protein structure, function and stability. EPL is a chemoselective ligation method that allows the selective ligation of unprotected polypeptides from synthetic and recombinant origin for the production of semi-synthetic protein samples of well-defined and homogeneous chemical composition. This method has been extensively used for the site-specific introduction of biophysical probes, unnatural amino acids, and increasingly complex post-translational modifications. Since it was introduced 10 years ago, EPL applications have grown increasingly more sophisticated in order to address even more complex biological questions. In this review, we highlight how this powerful technology combined with standard biochemical analysis techniques has been used to improve our ability to understand protein structure and function.
Berrade
Luis
L
Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, 1985 Zonal Avenue, PSC 616, Los Angeles, CA 90033, USA.
Camarero
Julio A
JA
D008956:Models, Chemical; D015394:Molecular Structure; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000235:genetics; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000138:chemical synthesis* / Q000737:chemistry / Q000235:genetics
D010455:Peptides; D011506:Proteins
J Pept Res. 1998 Apr;51(4):303-16
9560006
Gene. 1997 Jun 19;192(2):271-81
9224900
Biochem J. 1990 Jun 1;268(2):249-62
2163604
Adv Exp Med Biol. 2009;611:117-8
19400119
Angew Chem Int Ed Engl. 2004 Mar 5;43(11):1355-9
15368405
Science. 2003 Oct 24;302(5645):646-50
14576435
Org Lett. 2002 Jan 24;4(2):165-8
11796041
Cell. 2007 Feb 23;128(4):693-705
17320507
J Am Chem Soc. 2006 Oct 25;128(42):13840-6
17044712
Int J Biochem Cell Biol. 2007;39(12):2252-64
17693122
Biopolymers. 2007;88(2):308-24
17278124
J Org Chem. 2004 Jun 11;69(12):4145-51
15176841
J Mol Biol. 1999 Dec 17;294(5):1327-36
10600388
Gene. 1999 Apr 29;231(1-2):1-13
10231563
Angew Chem Int Ed Engl. 2003 Jan 27;42(4):431-4
12569509
FEBS Lett. 2008 Apr 2;582(7):1163-7
18331839
Angew Chem Int Ed Engl. 2006 Jan 30;45(6):973-6
16381044
Chemistry. 2001 Mar 2;7(5):1129-32
11303872
Biopolymers. 1999;51(5):333-42
10685044
Biochemistry. 2008 Sep 30;47(39):10407-19
18771288
Proc Natl Acad Sci U S A. 2003 Oct 14;100(21):12033-8
14530408
J Am Chem Soc. 2002 Aug 7;124(31):9113-20
12149015
Bioconjug Chem. 2009 Feb;20(2):360-6
19146394
Chem Biol. 2000 Apr;7(4):263-74
10780926
Curr Opin Chem Biol. 2005 Dec;9(6):561-9
16226484
Annu Rev Biochem. 1992;61:387-418
1497316
J Am Chem Soc. 2004 Nov 3;126(43):14004-12
15506763
Bioorg Med Chem. 2001 Sep;9(9):2479-84
11553489
J Am Chem Soc. 2004 Nov 17;126(45):14730-1
15535692
Bioorg Med Chem Lett. 2002 Aug 19;12(16):2079-83
12127508
Chem Biol. 1996 Dec;3(12):981-91
9000009
Chem Commun (Camb). 2008 Aug 28;(32):3723-5
18685756
Protein Sci. 1998 Nov;7(11):2256-64
9827992
Annu Rev Biochem. 2003;72:249-89
12626339
EMBO J. 2006 Jan 11;25(1):150-62
16362043
Mol Cell. 2005 Oct 28;20(2):251-62
16246727
J Biol Chem. 1998 Jun 26;273(26):16205-9
9632677
Proc Natl Acad Sci U S A. 1998 Jun 9;95(12):6705-10
9618476
Proc Natl Acad Sci U S A. 1999 Aug 31;96(18):10068-73
10468563
Biopolymers. 2008;90(3):450-8
17618518
Nucleic Acids Res. 1998 Nov 15;26(22):5109-15
9801307
Curr Protoc Protein Sci. 2001 May;Chapter 18:Unit18.4
18429140
Angew Chem Int Ed Engl. 1998 Feb 16;37(3):347-349
29711251
J Am Chem Soc. 2004 Dec 22;126(50):16368-78
15600338
Mol Cell. 2001 Sep;8(3):671-82
11583628
J Biol Chem. 2002 Mar 15;277(11):9512-9
11741964
Angew Chem Int Ed Engl. 2006 Oct 13;45(40):6622-46
17031879
Curr Opin Drug Discov Devel. 2006 Mar;9(2):251-60
16566295
Chembiochem. 2009 Mar 23;10(5):787-98
19199328
Chemistry. 2005 Apr 22;11(9):2756-72
15729676
J Am Chem Soc. 2004 Feb 4;126(4):1055-62
14746473
FEBS Lett. 1999 Oct 8;459(2):166-72
10518012
J Mol Biol. 2008 Jan 4;375(1):151-64
17936301
EMBO J. 2008 Sep 17;27(18):2444-56
18756270
J Am Chem Soc. 2006 Sep 6;128(35):11591-9
16939283
Angew Chem Int Ed Engl. 2008;47(52):10030-74
19072788
Chem Rec. 2006;6(4):183-93
16900466
Angew Chem Int Ed Engl. 2008;47(36):6851-5
18651678
Protein Sci. 1997 Jun;6(6):1129-38
9194173
Angew Chem Int Ed Engl. 2007;46(16):2814-8
17366504
Curr Opin Drug Discov Devel. 2002 Mar;5(2):251-60
11926131
Chembiochem. 2007 Aug 13;8(12):1363-6
17590879
Annu Rev Biochem. 2000;69:923-60
10966479
Chembiochem. 2007 Jan 2;8(1):32-6
17121405
Nat Methods. 2006 Jun;3(6):429-38
16721376
EMBO J. 2006 Jan 11;25(1):13-23
16395334
J Biol Chem. 2003 May 2;278(18):15744-8
12595522
J Am Chem Soc. 2005 Sep 21;127(37):12882-9
16159282
Biotechniques. 1999 Jul;27(1):110-4, 116, 118-20
10407673
Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13585-90
9811843
Science. 2006 Nov 10;314(5801):1004-7
17095703
Proc Natl Acad Sci U S A. 2004 Apr 27;101(17):6397-402
15087498
Chembiochem. 2007 Jun 18;8(9):1071-6
17508367
J Biol Chem. 2005 Sep 2;280(35):31019-26
15983037
Proc Natl Acad Sci U S A. 1989 Nov;86(21):8247-51
2682640
Acc Chem Res. 2009 Jan 20;42(1):107-16
18939858
Mol Cell. 2001 Dec;8(6):1277-89
11779503
Nature. 2008 Jun 5;453(7196):812-6
18449190
Mol Cell. 2004 Sep 10;15(5):813-23
15350224
Anal Biochem. 2003 Jun 15;317(2):226-32
12758261
J Am Chem Soc. 2008 Jul 16;130(28):9106-12
18570424
J Mol Biol. 2001 May 18;308(5):1045-62
11352590
Science. 1994 Nov 4;266(5186):776-9
7973629
J Mol Biol. 2005 Sep 2;351(5):939-47
16055150
Angew Chem Int Ed Engl. 2006 Feb 20;45(9):1408-12
16440394
J Biol Chem. 1999 Jun 25;274(26):18359-63
10373440
EMBO J. 1996 Oct 1;15(19):5146-53
8895558
Methods Enzymol. 2005;403:29-42
16473575
Protein Sci. 2008 Apr;17(4):736-47
18305193
J Am Chem Soc. 2002 Jul 31;124(30):8768-9
12137509
Chembiochem. 2006 Sep;7(9):1375-83
16897799
Genes Dev. 2002 Aug 1;16(15):1950-63
12154125
J Am Chem Soc. 2008 Jul 30;130(30):9702-7
18598026
J Am Chem Soc. 2007 Sep 19;129(37):11543-50
17715922
Proc Natl Acad Sci U S A. 2002 Jun 25;99(13):8536-41
12084914
Bioorg Med Chem Lett. 2005 May 16;15(10):2447-51
15863295
Curr Opin Chem Biol. 2006 Oct;10(5):487-91
16935551
Science. 2006 Feb 10;311(5762):844-7
16469925
Biochemistry. 2004 May 18;43(19):5698-706
15134444
J Biol Chem. 1999 Feb 12;274(7):3923-6
9933578
Chem Soc Rev. 2009 Feb;38(2):338-51
19169452
Proc Natl Acad Sci U S A. 1999 Jan 19;96(2):388-93
9892643
Angew Chem Int Ed Engl. 2002 Jun 17;41(12):2171-4
19746634
Angew Chem Int Ed Engl. 2006 Jun 26;45(26):4286-90
16739151
Bioorg Med Chem. 2006 Sep 15;14(18):6288-306
16725326
Protein Pept Lett. 2005 Nov;12(8):723-8
16305540
Proc Natl Acad Sci U S A. 1995 Dec 19;92(26):12485-9
8618926
Mol Cell. 2005 Jan 21;17(2):265-75
15664195
Protein Expr Purif. 2007 Aug;54(2):227-33
17493830