Systematic Engineering of a Protein Nanocage for High-Yield, Site-Specific Modification

2019 ◽  
Vol 141 (9) ◽  
pp. 3875-3884 ◽  
Author(s):  
Daniel D. Brauer ◽  
Emily C. Hartman ◽  
Daniel L. V. Bader ◽  
Zoe N. Merz ◽  
Danielle Tullman-Ercek ◽  
...  
Keyword(s):  
High Yield ◽  
Site Specific ◽  
Specific Modification

Systematic Engineering of a Protein Nanocage for High-Yield, Site-Specific Modification

2018 ◽  
Author(s):  
Daniel D. Brauer ◽  
Emily C. Hartman ◽  
Daniel L.V. Bader ◽  
Zoe N. Merz ◽  
Danielle Tullman-Ercek ◽  
...  
Keyword(s):  
Small Molecules ◽  
Protein Modification ◽  
Positive Charge ◽  
Specific Protein ◽  
High Yield ◽  
Site Specific ◽  
Protein Cage ◽  
Protein Carriers ◽  
Site Selective ◽  
Targeted Library

<div> <p>Site-specific protein modification is a widely-used strategy to attach drugs, imaging agents, or other useful small molecules to protein carriers. N-terminal modification is particularly useful as a high-yielding, site-selective modification strategy that can be compatible with a wide array of proteins. However, this modification strategy is incompatible with proteins with buried or sterically-hindered N termini, such as virus-like particles like the well-studied MS2 bacteriophage coat protein. To assess VLPs with improved compatibility with these techniques, we generated a targeted library based on the MS2-derived protein cage with N-terminal proline residues followed by three variable positions. We subjected the library to assembly, heat, and chemical selections, and we identified variants that were modified in high yield with no reduction in thermostability. Positive charge adjacent to the native N terminus is surprisingly beneficial for successful extension, and over 50% of the highest performing variants contained positive charge at this position. Taken together, these studies described nonintuitive design rules governing N-terminal extensions and identified successful extensions with high modification potential.</p> </div>

Site-specific modification of E. coli DNA polymerase I large fragment with pyridoxal 5'-phosphate

Biochemistry ◽  
1984 ◽  
Vol 23 (9) ◽  
pp. 2073-2078 ◽  
Author(s):  
Anup K. Hazra ◽  
Sevilla Detera-Wadleigh ◽  
Samuel H. Wilson
Keyword(s):  
Dna Polymerase ◽  
Large Fragment ◽  
Dna Polymerase I ◽  
Site Specific ◽  
E Coli ◽  
Specific Modification ◽  
Polymerase I

scholarly journals Site-specific N-glycosylation analysis of animal cell culture-derived Zika virus proteins

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alexander Pralow ◽  
Alexander Nikolay ◽  
Arnaud Leon ◽  
Yvonne Genzel ◽  
Erdmann Rapp ◽  
...  
Keyword(s):  
Zika Virus ◽  
Animal Cell ◽  
Gradient Centrifugation ◽  
Viral Envelope ◽  
Protein Glycosylation ◽  
High Yield ◽  
Structural Protein ◽  
Site Specific ◽  
E Protein ◽  
The Impact

AbstractHere, we present for the first time, a site-specific N-glycosylation analysis of proteins from a Brazilian Zika virus (ZIKV) strain. The virus was propagated with high yield in an embryo-derived stem cell line (EB66, Valneva SE), and concentrated by g-force step-gradient centrifugation. Subsequently, the sample was proteolytically digested with different enzymes, measured via a LC–MS/MS-based workflow, and analyzed in a semi-automated way using the in-house developed glyXtoolMS software. The viral non-structural protein 1 (NS1) was glycosylated exclusively with high-mannose structures on both potential N-glycosylation sites. In case of the viral envelope (E) protein, no specific N-glycans could be identified with this method. Nevertheless, N-glycosylation could be proved by enzymatic de-N-glycosylation with PNGase F, resulting in a strong MS-signal of the former glycopeptide with deamidated asparagine at the potential N-glycosylation site N444. This confirmed that this site of the ZIKV E protein is highly N-glycosylated but with very high micro-heterogeneity. Our study clearly demonstrates the progress made towards site-specific N-glycosylation analysis of viral proteins, i.e. for Brazilian ZIKV. It allows to better characterize viral isolates, and to monitor glycosylation of major antigens. The method established can be applied for detailed studies regarding the impact of protein glycosylation on antigenicity and human pathogenicity of many viruses including influenza virus, HIV and corona virus.

scholarly journals Dual, Site-Specific Modification of Antibodies by Using Solid-Phase Immobilized Microbial Transglutaminase

ChemBioChem ◽  
2017 ◽  
Vol 18 (19) ◽  
pp. 1923-1927 ◽  
Author(s):  
Philipp R. Spycher ◽  
Christian A. Amann ◽  
Jöri E. Wehrmüller ◽  
David R. Hurwitz ◽  
Olivier Kreis ◽  
...  
Keyword(s):  
Solid Phase ◽  
Microbial Transglutaminase ◽  
Site Specific ◽  
Specific Modification ◽  
Dual Site

High Yield, Site-Specific Coupling of N-Terminally Modified .beta.-Lactamase to a Proteolytically Derived Single-Sulfhydryl Murine Fab'

1994 ◽  
Vol 5 (6) ◽  
pp. 636-646 ◽  
Author(s):  
Stephen D. Mikolajczyk ◽  
Damon L. Meyer ◽  
James J. Starling ◽  
Kevin L. Law ◽  
Keith Rose ◽  
...  
Keyword(s):  
High Yield ◽  
Beta Lactamase ◽  
Site Specific

scholarly journals Expression of ΔF508 CFTR in normal mouse lung after site-specific modification of CFTR sequences by SFHR

Gene Therapy ◽  
2001 ◽  
Vol 8 (12) ◽  
pp. 961-965 ◽  
Author(s):  
KK Goncz ◽  
A Colosimo ◽  
B Dallapiccola ◽  
L Gagné ◽  
K Hong ◽  
...  
Keyword(s):  
Normal Mouse ◽  
Mouse Lung ◽  
Site Specific ◽  
Specific Modification ◽  
Δf508 Cftr

Methyl acetyl phosphate: A novel acetylating agent Its site-specific modification of human hemoglobin A

1986 ◽  
Vol 359 ◽  
pp. 193-201 ◽  
Author(s):  
Hiroshi Ueno ◽  
Maria A. Pospischil ◽  
Ronald Kluger ◽  
James M. Manning
Keyword(s):  
Human Hemoglobin ◽  
Acetyl Phosphate ◽  
Site Specific ◽  
Specific Modification ◽  
Hemoglobin A

scholarly journals Site-specific modification of the lactose operator with acetylaminofluorene

1983 ◽  
Vol 11 (15) ◽  
pp. 5093-5102 ◽  
Author(s):  
Gerhard Stöhrer ◽  
Judith A. Osband ◽  
Gabriel Alvarado-Urbina
Keyword(s):  
Site Specific ◽  
Specific Modification

scholarly journals Site specific modification of the human plasma proteome by methylglyoxal

2015 ◽  
Vol 289 (2) ◽  
pp. 155-162 ◽  
Author(s):  
Michael J. Kimzey ◽  
Owen R. Kinsky ◽  
Hussein N. Yassine ◽  
George Tsaprailis ◽  
Craig S. Stump ◽  
...  
Keyword(s):  
Human Plasma ◽  
Plasma Proteome ◽  
Site Specific ◽  
Specific Modification ◽  
Human Plasma Proteome
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