The scope of phage display for membrane proteins.
Numerous examples of phage display applied to soluble proteins demonstrate the power of the technique for protein engineering, affinity reagent discovery and structure-function studies. Recent reports have expanded phage display to include membrane proteins (MPs). The scope and limitations of MP display remain undefined. Therefore, we report data from the phage display of representative types of membrane-associated proteins including plasma, nuclear, peripheral, single and multipass. The peripheral MP neuromodulin displays robustly with packaging by conventional M13-KO7 helper phage. The monotopic MP Nogo-66 can also display on the phage surface, if packaged by the modified M13-KO7(+) helper phage. The modified phage coat of KO7(+) can better mimic the zwitterionic character of the plasma membrane. Four examples of putatively α-helical, integral MPs failed to express as fusions to an anchoring phage coat protein and therefore did not display on the phage surface. However, the β-barrel MPs ShuA (Shigella heme uptake A) and MOMP (major outer membrane protein), which pass through the membrane 22 and 16 times, respectively, can display surprisingly well on the surfaces of both conventional and KO7(+) phages. The results provide a guide for protein engineering and large-scale mutagenesis enabled by the phage display of MPs.
Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697, USA.
Rosemarie
R
Vithayathil
Richard M
RM
Hooy
Melanie J
MJ
Cocco
Gregory A
GA
Weiss
D017104:Bacteriophage M13 / Q000235:genetics*; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular / Q000379:methods; D005821:Genetic Techniques; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism*; D008958:Models, Molecular; D016296:Mutagenesis / Q000235:genetics; D019151:Peptide Library; D015202:Protein Engineering / Q000379:methods; D017433:Protein Structure, Secondary / Q000235:genetics
D036022:Capsid Proteins; D008565:Membrane Proteins; D019151:Peptide Library; C025668:coat protein, Bacteriophage M13