publication . Article . Other literature type . 2014

Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications

Christian Tiede; Anna Tang; Robin Owen; Sarah Deacon; Ray Owens; Darren Tomlinson; Joanne Nettleship; Michael McPherson;
Open Access
  • Published: 25 Mar 2014 Journal: Protein Engineering, Design and Selection, volume 27, pages 145-155 (issn: 1741-0126, eissn: 1741-0134, Copyright policy)
  • Publisher: Oxford University Press (OUP)
  • Country: United Kingdom
Abstract
None: We have designed a novel non-antibody scaffold protein, termed Adhiron, based on a phytocystatin consensus sequence. The Adhiron scaffold shows high thermal stability (Tm ca. 101°C), and is expressed well in Escherichia coli. We have determined the X-ray crystal structure of the Adhiron scaffold to 1.75 Å resolution revealing a compact cystatin-like fold. We have constructed a phage-display library in this scaffold by insertion of two variable peptide regions. The library is of high quality and complexity comprising 1.3 × 10(10) clones. To demonstrate library efficacy, we screened against the yeast Small Ubiquitin-like Modifier (SUMO). In selected clones, ...
Subjects
free text keywords: Biotechnology, Biochemistry, Bioengineering, Molecular Biology, Original Articles, consensus protein, high specificity binding, non-antibody-binding protein, protein–protein interaction, SUMO, Q1, R1, RM
51 references, page 1 of 4

Afonine,P.V., Grosse-Kunstleve,R.W., Echols,N., et al. (2012) Acta Crystallogr. D, 68, 352 - 367.

Binz,H.K., Stumpp,M.T., Forrer,P., Amstutz,P. and Pluckthun,A. (2003) J. Mol. Biol., 332, 489 - 503.

Bode,W., Engh,R., Musil,D., Thiele,U., Huber,R., Karshikov,A., Brzin,J., Kos,J. and Turk,V. (1988) EMBO J., 7, 2593 - 2599.

Carter,P.J. (2011) Exp. Cell Res., 317, 1261- 1269.

Colwill,K., Graslund,S. and Renewable Prot Binder Working,G. (2011) Nat. Methods, 8, 551 - 558.

Corpet,F. (1988) Nucleic Acids Res, 16, 10881 - 10890.

Emsley,P., Lohkamp,B., Scott,W.G. and Cowtan,K. (2010) Acta Crystallogr. D, 66, 486 - 501.

Evans,P.R. (2006) Acta Crystallogr. D, 62, 72 - 82.

Forrer,P., Binz,H.K., Stumpp,M.T. and Pluckthun,A. (2004) Chembiochem, 5, 183 - 189.

Gebauer,M. and Skerra,A. (2009) Curr. Opin. Chem. Biol., 13, 245 - 255.

Gilbreth,R.N., Truong,K., Madu,I., Koide,A., Wojcik,J.B., Li,N.-S., Piccirilli,J.A., Chen,Y. and Koide,S. (2011) Proc. Natl. Acad. Sci. USA, 108, 7751- 7756.

Grebien,F., Hantschel,O., Wojcik,J., et al. (2011) Cell, 147, 306 - 319.

Hoogenboom,H.R., Griffiths,A.D., Johnson,K.S., Chiswell,D.J., Hudson,P. and Winter,G. (1991) Nucleic Acids Res., 19, 4133- 4137.

Horton,R., Cai,Z., Ho,S. and Pease,L. (1990) Biotechniques, 8, 528 - 535.

Irene,D., Chung,T.-Y., Chen,B.-J., Liu,T.-H., Li,F.-Y., Tzen,J.T.C., Wang,C.-I. and Chyan,C.L. (2012) PLoS ONE, 7, e47865.

51 references, page 1 of 4
Abstract
None: We have designed a novel non-antibody scaffold protein, termed Adhiron, based on a phytocystatin consensus sequence. The Adhiron scaffold shows high thermal stability (Tm ca. 101°C), and is expressed well in Escherichia coli. We have determined the X-ray crystal structure of the Adhiron scaffold to 1.75 Å resolution revealing a compact cystatin-like fold. We have constructed a phage-display library in this scaffold by insertion of two variable peptide regions. The library is of high quality and complexity comprising 1.3 × 10(10) clones. To demonstrate library efficacy, we screened against the yeast Small Ubiquitin-like Modifier (SUMO). In selected clones, ...
Subjects
free text keywords: Biotechnology, Biochemistry, Bioengineering, Molecular Biology, Original Articles, consensus protein, high specificity binding, non-antibody-binding protein, protein–protein interaction, SUMO, Q1, R1, RM
51 references, page 1 of 4

Afonine,P.V., Grosse-Kunstleve,R.W., Echols,N., et al. (2012) Acta Crystallogr. D, 68, 352 - 367.

Binz,H.K., Stumpp,M.T., Forrer,P., Amstutz,P. and Pluckthun,A. (2003) J. Mol. Biol., 332, 489 - 503.

Bode,W., Engh,R., Musil,D., Thiele,U., Huber,R., Karshikov,A., Brzin,J., Kos,J. and Turk,V. (1988) EMBO J., 7, 2593 - 2599.

Carter,P.J. (2011) Exp. Cell Res., 317, 1261- 1269.

Colwill,K., Graslund,S. and Renewable Prot Binder Working,G. (2011) Nat. Methods, 8, 551 - 558.

Corpet,F. (1988) Nucleic Acids Res, 16, 10881 - 10890.

Emsley,P., Lohkamp,B., Scott,W.G. and Cowtan,K. (2010) Acta Crystallogr. D, 66, 486 - 501.

Evans,P.R. (2006) Acta Crystallogr. D, 62, 72 - 82.

Forrer,P., Binz,H.K., Stumpp,M.T. and Pluckthun,A. (2004) Chembiochem, 5, 183 - 189.

Gebauer,M. and Skerra,A. (2009) Curr. Opin. Chem. Biol., 13, 245 - 255.

Gilbreth,R.N., Truong,K., Madu,I., Koide,A., Wojcik,J.B., Li,N.-S., Piccirilli,J.A., Chen,Y. and Koide,S. (2011) Proc. Natl. Acad. Sci. USA, 108, 7751- 7756.

Grebien,F., Hantschel,O., Wojcik,J., et al. (2011) Cell, 147, 306 - 319.

Hoogenboom,H.R., Griffiths,A.D., Johnson,K.S., Chiswell,D.J., Hudson,P. and Winter,G. (1991) Nucleic Acids Res., 19, 4133- 4137.

Horton,R., Cai,Z., Ho,S. and Pease,L. (1990) Biotechniques, 8, 528 - 535.

Irene,D., Chung,T.-Y., Chen,B.-J., Liu,T.-H., Li,F.-Y., Tzen,J.T.C., Wang,C.-I. and Chyan,C.L. (2012) PLoS ONE, 7, e47865.

51 references, page 1 of 4
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