Abstract
Phosphopantetheinyl transferases (PPTase) Sfp and AcpS catalyze a highly efficient reaction that conjugates chemical probes of diverse structures to proteins. PPTases have been widely used for site-specific protein labeling and live cell imaging of the target proteins. Here we describe the use of PPTase-catalyzed protein labeling in protein engineering by facilitating high-throughput phage selection.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Yin J, Lin AJ, Golan DE, Walsh CT (2006) Site-specific protein labeling by Sfp phosphopantetheinyl transferase. Nat Protoc 1:280–285
Lambalot RH, Gehring AM, Flugel RS, Zuber P, LaCelle M, Marahiel MA, Reid R, Khosla C, Walsh CT (1996) A new enzyme superfamily—the phosphopantetheinyl transferases. Chem Biol 3:923–936
La Clair JJ, Foley TL, Schegg TR, Regan CM, Burkart MD (2004) Manipulation of carrier proteins in antibiotic biosynthesis. Chem Biol 11:195–201
Yin J, Liu F, Li X, Walsh CT (2004) Labeling proteins with small molecules by site-specific posttranslational modification. J Am Chem Soc 126:7754–7755
Yin J, Liu F, Schinke M, Daly C, Walsh CT (2004) Phagemid encoded small molecules for high throughput screening of chemical libraries. J Am Chem Soc 126:13570–13571
Yin J, Lin AJ, Buckett PD, Wessling-Resnick M, Golan DE, Walsh CT (2005) Single-cell FRET imaging of transferrin receptor trafficking dynamics by Sfp-catalyzed, site-specific protein labeling. Chem Biol 12:999–1006
Yin J, Straight PD, Hrvatin S, Dorrestein PC, Bumpus SB, Jao C, Kelleher NL, Kolter R, Walsh CT (2007) Genome-wide high-throughput mining of natural-product biosynthetic gene clusters by phage display. Chem Biol 14:303–312
Zhou Z, Cironi P, Lin AJ, Xu Y, Hrvatin S, Golan DE, Silver PA, Walsh CT, Yin J (2007) Genetically encoded short peptide tags for orthogonal protein labeling by Sfp and AcpS phosphopantetheinyl transferases. ACS Chem Biol 2:337–346
Yin J, Straight PD, McLoughlin SM, Zhou Z, Lin AJ, Golan DE, Kelleher NL, Kolter R, Walsh CT (2005) Genetically encoded short peptide tag for versatile protein labeling by Sfp phosphopantetheinyl transferase. Proc Natl Acad Sci U S A 102:15815–15820
George N, Pick H, Vogel H, Johnsson N, Johnsson K (2004) Specific labeling of cell surface proteins with chemically diverse compounds. J Am Chem Soc 126:8896–8897
Waichman S, Bhagawati M, Podoplelova Y, Reichel A, Brunk A, Paterok D, Piehler J (2010) Functional immobilization and patterning of proteins by an enzymatic transfer reaction. Anal Chem 82:1478–1485
Wong LS, Okrasa K, Micklefield J (2010) Site-selective immobilisation of functional enzymes on to polystyrene nanoparticles. Org Biomol Chem 8:782–787
Wong LS, Thirlway J, Micklefield J (2008) Direct site-selective covalent protein immobilization catalyzed by a phosphopantetheinyl transferase. J Am Chem Soc 130:12456–12464
Band PA, Blais S, Neubert TA, Cardozo TJ, Ichtchenko K (2010) Recombinant derivatives of botulinum neurotoxin A engineered for trafficking studies and neuronal delivery. Protein Expr Purif 71:62–73
Kawamura T, Stephens B, Qin L, Yin X, Dores MR, Smith TH, Grimsey N, Abagyan R, Trejo J, Kufareva I, Fuster MM, Salanga CL, Handel TM (2014) A general method for site specific fluorescent labeling of recombinant chemokines. PLoS One 9:e81454
Wang Y, Zhou Z, Walsh CT, McMahon AP (2009) Selective translocation of intracellular Smoothened to the primary cilium in response to Hedgehog pathway modulation. Proc Natl Acad Sci U S A 106:2623–2628
Zhao B, Bhuripanyo K, Schneider J, Zhang K, Schindelin H, Boone D, Yin J (2012) Specificity of the E1-E2-E3 enzymatic cascade for ubiquitin C-terminal sequences identified by phage display. ACS Chem Biol 7:2027–2035
Zhao B, Bhuripanyo K, Zhang K, Kiyokawa H, Schindelin H, Yin J (2012) Orthogonal ubiquitin transfer through engineered E1-E2 cascades for protein ubiquitination. Chem Biol 19:1265–1277
Zhang K, Li H, Bhuripanyo K, Zhao B, Chen TF, Zheng N, Yin J (2013) Engineering new protein-protein interactions on the beta-propeller fold by yeast cell surface display. Chembiochem 14:426–430
Lee I, Schindelin H (2008) Structural insights into E1-catalyzed ubiquitin activation and transfer to conjugating enzymes. Cell 134:268–278
Schulman BA, Harper JW (2009) Ubiquitin-like protein activation by E1 enzymes: the apex for downstream signalling pathways. Nat Rev Mol Cell Biol 10:319–331
Acknowledgements
This work was supported by a National Science Foundation CAREER award (1057092) and a National Institute of Health grant 1R01GM104498 to J.Y.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Zhao, B. et al. (2015). Phage Selection Assisted by Sfp Phosphopantetheinyl Transferase-Catalyzed Site-Specific Protein Labeling. In: Gautier, A., Hinner, M. (eds) Site-Specific Protein Labeling. Methods in Molecular Biology, vol 1266. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2272-7_11
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2272-7_11
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2271-0
Online ISBN: 978-1-4939-2272-7
eBook Packages: Springer Protocols