Covalent Surface Modification of Prions: A Mass Spectrometry-Based Means of Detecting Distinctive Structural Features of Prion Strains

Biochemistry ◽  
2016 ◽  
Vol 55 (6) ◽  
pp. 894-902 ◽  
Author(s):  
Christopher J. Silva ◽  
Melissa L. Erickson-Beltran ◽  
Irina C. Dynin
Keyword(s):  
Mass Spectrometry ◽  
Surface Modification ◽  
Structural Features ◽  
Prion Strains

Rapid Online Buffer Exchange: A Method for Screening of Proteins, Protein Complexes, and Cell Lysates by Native Mass Spectrometry

2019 ◽  
Author(s):  
Zachary VanAernum ◽  
Florian Busch ◽  
Benjamin J. Jones ◽  
Mengxuan Jia ◽  
Zibo Chen ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Speed ◽  
Structural Information ◽  
Protein Complexes ◽  
High Sensitivity ◽  
Native Mass Spectrometry ◽  
Structural Features ◽  
Consumer Products ◽  
Cell Lysates ◽  
Protein Expression And Purification

It is important to assess the identity and purity of proteins and protein complexes during and after protein purification to ensure that samples are of sufficient quality for further biochemical and structural characterization, as well as for use in consumer products, chemical processes, and therapeutics. Native mass spectrometry (nMS) has become an important tool in protein analysis due to its ability to retain non-covalent interactions during measurements, making it possible to obtain protein structural information with high sensitivity and at high speed. Interferences from the presence of non-volatiles are typically alleviated by offline buffer exchange, which is timeconsuming and difficult to automate. We provide a protocol for rapid online buffer exchange (OBE) nMS to directly screen structural features of pre-purified proteins, protein complexes, or clarified cell lysates. Information obtained by OBE nMS can be used for fast (<5 min) quality control and can further guide protein expression and purification optimization.

Using Mass Spectrometry to Investigate the Structural Features of Photocrosslinked Co-Networks based on Gelatin and Poly(ethylene glycol) Methacrylates

2012 ◽  
Vol 1403 ◽  
Author(s):  
Benjamin F. Pierce ◽  
Axel T. Neffe ◽  
Andreas Lendlein
Keyword(s):  
Mass Spectrometry ◽  
Ethylene Glycol ◽  
Degradation Products ◽  
Structural Features ◽  
Valuable Insight ◽  
Poly Ethylene Glycol ◽  
Buffer Solution ◽  
Molecular Weight Range ◽  
Poly Ethylene

ABSTRACTGelatin was functionalized with glycidyl methacrylate and photocrosslinked in the presence of poly(ethylene glycol) dimethacrylate (PEGDMA) or poly(ethylene glycol) monomethacrylate (PEGMA) to create a biopolymer-based system with tailorable properties. These co-networks were hydrolyzed using 6 M HCl and the degradation products were analyzed and identified using matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. This technique successfully identified gelatin-derived peptides such as FLPEPPE, SFLPEPPE, and SFLPEPPEE as well as an accompanying PEG-g-poly(methacrylic acid) component. No oligo- or polymethacrylates were monitored at any molecular weight range above m/z = 500, which indicated that they possessed lower molecular weights. An in vitro hydrolytic degradation experiment performed in pH 7.4 PBS buffer solution at 37 °C showed that these networks, which were prepared without the addition of a potentially toxic photoinitiator, exhibited mass loss of up to 50 wt% at 6 weeks of incubation time. These results provide valuable insight into how these functional gelatin-based co-network biomaterials will perform in a biological setting.

scholarly journals Analysis of the composition of thermolysis products of ash-free concentrates obtained on the basis of oil shale of the Ayuvinskoye deposit (Komi Republic)

2021 ◽  
Vol 10 ◽  
pp. 33-41
Author(s):  
N. S. Burdelnaya ◽  
◽  
D. A. Bushnev ◽  
I. N. Burtsev ◽  
D. V. Kuzmin ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Organic Matter ◽  
Oil Shale ◽  
Fossil Fuels ◽  
Sedimentary Rocks ◽  
Structural Features ◽  
Komi Republic ◽  
Gas Chromatography Mass Spectrometry ◽  
Alkyl Chains

The treatment with N-methylpyrrolidone of sedimentary rocks of the Ayuvinskoye deposit allowed obtaining ash-free concentrates with different yields, depending on the Corg content in the rock. The structure of the resulting concentrates was studied by elemental analysis, thermogravimetry, and pyrolysis, followed by analysis of the products by gas chromatography-mass spectrometry. The TGA curves indicated similar structural features of the organic matter of the rocks and the concentrates obtained from them. The composition of the thermolysis products of the concentrate indicated the preferential extraction of aliphatic structures, represented by n-alkyl chains, relative to aromatic fragments, which was associated with the specific structure of the initial organic matter of solid fossil fuels.

scholarly journals Rapid Online Buffer Exchange: A Method for Screening of Proteins, Protein Complexes, and Cell Lysates by Native Mass Spectrometry

2019 ◽  
Author(s):  
Zachary VanAernum ◽  
Florian Busch ◽  
Benjamin J. Jones ◽  
Mengxuan Jia ◽  
Zibo Chen ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Speed ◽  
Structural Information ◽  
Protein Complexes ◽  
High Sensitivity ◽  
Native Mass Spectrometry ◽  
Structural Features ◽  
Consumer Products ◽  
Cell Lysates ◽  
Protein Expression And Purification

It is important to assess the identity and purity of proteins and protein complexes during and after protein purification to ensure that samples are of sufficient quality for further biochemical and structural characterization, as well as for use in consumer products, chemical processes, and therapeutics. Native mass spectrometry (nMS) has become an important tool in protein analysis due to its ability to retain non-covalent interactions during measurements, making it possible to obtain protein structural information with high sensitivity and at high speed. Interferences from the presence of non-volatiles are typically alleviated by offline buffer exchange, which is timeconsuming and difficult to automate. We provide a protocol for rapid online buffer exchange (OBE) nMS to directly screen structural features of pre-purified proteins, protein complexes, or clarified cell lysates. Information obtained by OBE nMS can be used for fast (<5 min) quality control and can further guide protein expression and purification optimization.

scholarly journals Modulation of Natural HLA-B*27:05 Ligandome by Ankylosing Spondylitis-associated Endoplasmic Reticulum Aminopeptidase 2 (ERAP2)

2020 ◽  
Vol 19 (6) ◽  
pp. 994-1004 ◽  
Author(s):  
Elena Lorente ◽  
Miguel G. Fontela ◽  
Eilon Barnea ◽  
Antonio J. Martín-Galiano ◽  
Carmen Mir ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Endoplasmic Reticulum ◽  
Bioinformatics Analysis ◽  
Structural Features ◽  
Live Cells ◽  
Pathogenic Role ◽  
Antigen Binding Site ◽  
Cell Clones ◽  
Hydrophobicity Profile

The HLA-B*27:05 allele and the endoplasmic reticulum-resident aminopeptidases are strongly associated with AS, a chronic inflammatory spondyloarthropathy. This study examined the effect of ERAP2 in the generation of the natural HLA-B*27:05 ligandome in live cells. Complexes of HLA-B*27:05-bound peptide pools were isolated from human ERAP2-edited cell clones, and the peptides were identified using high-throughput mass spectrometry analyses. The relative abundance of a thousand ligands was established by quantitative tandem mass spectrometry and bioinformatics analysis. The residue frequencies at different peptide position, identified in the presence or absence of ERAP2, determined structural features of ligands and their interactions with specific pockets of the antigen-binding site of the HLA-B*27:05 molecule. Sequence alignment of ligands identified with species of bacteria associated with HLA-B*27-dependent reactive arthritis was performed. In the absence of ERAP2, peptides with N-terminal basic residues and minority canonical P2 residues are enriched in the natural ligandome. Further, alterations of residue frequencies and hydrophobicity profile at P3, P7, and PΩ positions were detected. In addition, several ERAP2-dependent cellular peptides were highly similar to protein sequences of arthritogenic bacteria, including one human HLA-B*27:05 ligand fully conserved in a protein from Campylobacter jejuni. These findings highlight the pathogenic role of this aminopeptidase in the triggering of AS autoimmune disease.

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