scholarly journals Mutational analysis of the major coat protein of M13 identifies residues that control protein display

2008 ◽  
Vol 9 (4) ◽  
pp. 647-654 ◽  
Author(s):  
Gregory A. Weiss ◽  
James A. Wells ◽  
Sachdev S. Sidhu
Keyword(s):  
Coat Protein ◽  
Mutational Analysis ◽  
Major Coat Protein ◽  
Control Protein

scholarly journals Uniqueness of RNA Coliphage Qβ Display System in Directed Evolutionary Biotechnology

Viruses ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 568
Author(s):  
Godwin W. Nchinda ◽  
Nadia Al-Atoom ◽  
Mamie T. Coats ◽  
Jacqueline M. Cameron ◽  
Alain Bopda Waffo
Keyword(s):  
Life Cycle ◽  
Coat Protein ◽  
Phage Display ◽  
Display System ◽  
Phage Display Technology ◽  
Library Size ◽  
Exterior Surface ◽  
Major Coat Protein ◽  
Display Technology ◽  
Minor Coat Protein

Phage display technology involves the surface genetic engineering of phages to expose desirable proteins or peptides whose gene sequences are packaged within phage genomes, thereby rendering direct linkage between genotype with phenotype feasible. This has resulted in phage display systems becoming invaluable components of directed evolutionary biotechnology. The M13 is a DNA phage display system which dominates this technology and usually involves selected proteins or peptides being displayed through surface engineering of its minor coat proteins. The displayed protein or peptide’s functionality is often highly reduced due to harsh treatment of M13 variants. Recently, we developed a novel phage display system using the coliphage Qβ as a nano-biotechnology platform. The coliphage Qβ is an RNA phage belonging to the family of Leviviridae, a long investigated virus. Qβ phages exist as a quasispecies and possess features making them comparatively more suitable and unique for directed evolutionary biotechnology. As a quasispecies, Qβ benefits from the promiscuity of its RNA dependent RNA polymerase replicase, which lacks proofreading activity, and thereby permits rapid variant generation, mutation, and adaptation. The minor coat protein of Qβ is the readthrough protein, A1. It shares the same initiation codon with the major coat protein and is produced each time the ribosome translates the UGA stop codon of the major coat protein with the of misincorporation of tryptophan. This misincorporation occurs at a low level (1/15). Per convention and definition, A1 is the target for display technology, as this minor coat protein does not play a role in initiating the life cycle of Qβ phage like the pIII of M13. The maturation protein A2 of Qβ initiates the life cycle by binding to the pilus of the F+ host bacteria. The extension of the A1 protein with a foreign peptide probe recognizes and binds to the target freely, while the A2 initiates the infection. This avoids any disturbance of the complex and the necessity for acidic elution and neutralization prior to infection. The combined use of both the A1 and A2 proteins of Qβ in this display system allows for novel bio-panning, in vitro maturation, and evolution. Additionally, methods for large library size construction have been improved with our directed evolutionary phage display system. This novel phage display technology allows 12 copies of a specific desired peptide to be displayed on the exterior surface of Qβ in uniform distribution at the corners of the phage icosahedron. Through the recently optimized subtractive bio-panning strategy, fusion probes containing up to 80 amino acids altogether with linkers, can be displayed for target selection. Thus, combined uniqueness of its genome, structure, and proteins make the Qβ phage a desirable suitable innovation applicable in affinity maturation and directed evolutionary biotechnology. The evolutionary adaptability of the Qβ phage display strategy is still in its infancy. However, it has the potential to evolve functional domains of the desirable proteins, glycoproteins, and lipoproteins, rendering them superior to their natural counterparts.

Local Dynamics of the M13 Major Coat Protein in Different Membrane-Mimicking Systems†

Biochemistry ◽  
1996 ◽  
Vol 35 (48) ◽  
pp. 15467-15473 ◽  
Author(s):  
David Stopar ◽  
Ruud B. Spruijt ◽  
Cor J. A. M. Wolfs ◽  
Marcus A. Hemminga
Keyword(s):  
Coat Protein ◽  
Major Coat Protein ◽  
Local Dynamics

scholarly journals A Survey of Resistance to Tomato bushy stunt virus in the Genus Nicotiana Reveals That the Hypersensitive Response Is Triggered by One of Three Different Viral Proteins

2013 ◽  
Vol 26 (2) ◽  
pp. 240-248 ◽  
Author(s):  
Carlos A. Angel ◽  
James E. Schoelz
Keyword(s):  
Coat Protein ◽  
Hypersensitive Response ◽  
Rna Silencing ◽  
Mutational Analysis ◽  
Movement Protein ◽  
Viral Proteins ◽  
Tomato Bushy Stunt Virus ◽  
Necrosis Virus ◽  
Nicotiana Glutinosa ◽  
Cucumber Necrosis Virus

In this study, we screened 22 Nicotiana spp. for resistance to the tombusviruses Tomato bushy stunt virus (TBSV), Cucumber necrosis virus, and Cymbidium ringspot virus. Eighteen species were resistant, and resistance was manifested in at least two different categories. In all, 13 species responded with a hypersensitive response (HR)-type resistance, whereas another five were resistant but either had no visible response or responded with chlorotic lesions rather than necrotic lesions. Three different TBSV proteins were found to trigger HR in Nicotiana spp. in an agroinfiltration assay. The most common avirulence (avr) determinant was the TBSV coat protein P41, a protein that had not been previously recognized as an avr determinant. A mutational analysis confirmed that the coat protein rather than the viral RNA sequence was responsible for triggering HR, and it triggered HR in six species in the Alatae section. The TBSV P22 movement protein triggered HR in two species in section Undulatae (Nicotiana glutinosa and N. edwardsonii) and one species in section Alatae (N. forgetiana). The TBSV P19 RNA silencing suppressor protein triggered HR in sections Sylvestres (N. sylvestris), Nicotiana (N. tabacum), and Alatae (N. bonariensis). In general, Nicotiana spp. were capable of recognizing only one tombusvirus avirulence determinant, with the exceptions of N. bonariensis and N. forgetiana, which were each able to recognize P41, as well as P19 and P22, respectively. Agroinfiltration failed to detect the TBSV avr determinants responsible for triggering HR in N. arentsii, N. undulata, and N. rustica. This study illustrates the breadth and variety of resistance responses to tombusviruses that exists in the Nicotiana genus.

scholarly journals Major coat protein and single-stranded DNA-binding protein of filamentous virus Pf3.

1984 ◽  
Vol 81 (3) ◽  
pp. 699-703 ◽  
Author(s):  
D. G. Putterman ◽  
A. Casadevall ◽  
P. D. Boyle ◽  
H. L. Yang ◽  
B. Frangione ◽  
...  
Keyword(s):  
Coat Protein ◽  
Dna Binding ◽  
Binding Protein ◽  
Dna Binding Protein ◽  
Single Stranded Dna ◽  
Major Coat Protein

Reconstitution of the M13 Major Coat Protein and Its Transmembrane Peptide Segment on a DNA Template†

Biochemistry ◽  
2007 ◽  
Vol 46 (29) ◽  
pp. 8579-8591 ◽  
Author(s):  
Weijun Li ◽  
Itai Suez ◽  
Francis C. Szoka
Keyword(s):  
Coat Protein ◽  
Major Coat Protein ◽  
Dna Template

Accessibility and dynamics of Cys Residues in bacteriophage IKe and M13 major coat protein mutants

Biochemistry ◽  
1995 ◽  
Vol 34 (38) ◽  
pp. 12388-12397 ◽  
Author(s):  
Amir R. Khan ◽  
Karen A. Williams ◽  
Joan M. Boggs ◽  
Charles M. Deber
Keyword(s):  
Coat Protein ◽  
Major Coat Protein ◽  
Protein Mutants
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