Chemical cross-linking with a diazirine photoactivatable cross-linker investigated by MALDI- and ESI-MS/MS

2010 ◽  
Vol 45 (8) ◽  
pp. 892-899 ◽  
Author(s):  
Alexandre F. Gomes ◽  
Fabio C. Gozzo
Keyword(s):  
Cross Linking ◽  
Esi Ms ◽  
Cross Linker ◽  
Chemical Cross Linking

scholarly journals Influence of Cross-Linker Polarity on Selectivity towards Lysine Side Chains

2020 ◽  
Author(s):  
Jan Fiala ◽  
Zdeněk Kukačka ◽  
Petr Novák
Keyword(s):  
Structural Information ◽  
Protein Complexes ◽  
Water Soluble ◽  
Cross Linking ◽  
Side Chains ◽  
Model Protein ◽  
Cross Linker ◽  
Progressive Technology ◽  
The Impact ◽  
Chemical Cross Linking

The combination of chemical cross-linking and mass spectrometry is currently a progressive technology for deriving structural information of proteins and protein complexes. In addition, chemical cross-linking is a powerful tool for stabilizing macromolecular complexes for single particle cryo-electron microscopy. Broad pallets of cross-linking chemistry, currently available for the majority of cross-linking experiments, still rely on the amine-reactive N-hydroxysuccinimide esters targeting mainly N-termini and lysine side chains. These cross-linkers are divided into two groups: water soluble and water insoluble; and research teams prefer one or another speculating on the benefits of their choice. However, the effect of cross-linker polarity on the outcome of cross-linking reaction has never been studied. Herein, we use both polar (bis(sulfosuccinimidyl) glutarate) and non-polar (disuccinimidyl glutarate) cross-linkers and systematically investigated the impact of cross-linker hydrophobicity on resulting distance constraints, using bovine serum albumin as a model protein.

Detection of a Myc-associated protein by chemical cross-linking

1989 ◽  
Vol 9 (2) ◽  
pp. 865-868
Author(s):  
D A Gillespie ◽  
R N Eisenman
Keyword(s):  
Molecular Weight ◽  
Half Life ◽  
Nuclear Protein ◽  
Cross Linking ◽  
Cross Linker ◽  
Chemical Cross Linking

A single nuclear protein (Myc-associated protein) can be specifically cross-linked to avian Myc proteins by treatment of nuclei or cells with the reversible cross-linker dimethyl 3,3'-dithiobis-propionimidate. Myc-associated protein has a molecular weight of approximately 500,000, is not detectably phosphorylated and, in contrast to Myc, has a long apparent half-life of greater than 3 h.

scholarly journals Gelatin-Based Hydrogels through Homobifunctional Triazolinediones Targeting Tyrosine Residues

Molecules ◽  
2019 ◽  
Vol 24 (3) ◽  
pp. 589 ◽  
Author(s):  
Roberto Guizzardi ◽  
Luca Vaghi ◽  
Marcello Marelli ◽  
Antonino Natalello ◽  
Ivan Andreosso ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Drug Delivery Systems ◽  
Chemical Nature ◽  
Mechanical Stability ◽  
Cross Linking ◽  
Swelling Properties ◽  
Tyrosine Residues ◽  
Cross Linker ◽  
The Cross ◽  
Chemical Cross Linking

Gelatin is a biopolymer with interesting properties that can be useful for biomaterial design for different applications such as drug delivery systems, or 3D scaffolds for tissue engineering. However, gelatin suffers from poor mechanical stability at physiological temperature, hence methods for improving its properties are highly desirable. In the present work, a new chemical cross-linking strategy based on triazolinedione ene-type chemistry towards stable hydrogel is proposed. Two different homobifunctional 1,2,4-triazoline-3,5(4H)-diones, namely 4,4′-hexane-1,6-diylbis(3H-1,2,4-triazoline-3,5(4H)-dione) 1 and 4,4′-[methylenebis(4,1-phenylene)]bis(3H-1,2,4-triazoline-3,5(4H)-dione) 2 were used as cross-linkers in different ratio to tyrosine residues in gelatin. The reaction was proved effective in all experimented conditions and hydrogels featured with different thermal stability were obtained. In general, the higher the cross-linker/tyrosine ratio, the more thermostable the hydrogel. The swelling properties are strictly dependent upon the chemical nature of the cross-linker.

scholarly journals Detection of a Myc-associated protein by chemical cross-linking.

1989 ◽  
Vol 9 (2) ◽  
pp. 865-868 ◽  
Author(s):  
D A Gillespie ◽  
R N Eisenman
Keyword(s):  
Molecular Weight ◽  
Half Life ◽  
Nuclear Protein ◽  
Cross Linking ◽  
Cross Linker ◽  
Chemical Cross Linking

A single nuclear protein (Myc-associated protein) can be specifically cross-linked to avian Myc proteins by treatment of nuclei or cells with the reversible cross-linker dimethyl 3,3'-dithiobis-propionimidate. Myc-associated protein has a molecular weight of approximately 500,000, is not detectably phosphorylated and, in contrast to Myc, has a long apparent half-life of greater than 3 h.

scholarly journals Proximity-enhanced SuFEx chemical cross-linker for specific and multitargeting cross-linking mass spectrometry

2018 ◽  
Vol 115 (44) ◽  
pp. 11162-11167 ◽  
Author(s):  
Bing Yang ◽  
Haifan Wu ◽  
Paul D. Schnier ◽  
Yansheng Liu ◽  
Jun Liu ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Current Method ◽  
Thermal Fluctuations ◽  
Limited Range ◽  
Live Cells ◽  
Cross Linking ◽  
Cross Links ◽  
Cross Linker ◽  
Sulfonyl Fluoride ◽  
Chemical Cross Linking

Chemical cross-linking mass spectrometry (CXMS) is being increasingly used to study protein assemblies and complex protein interaction networks. Existing CXMS chemical cross-linkers target only Lys, Cys, Glu, and Asp residues, limiting the information measurable. Here we report a “plant-and-cast” cross-linking strategy that employs a heterobifunctional cross-linker that contains a highly reactive succinimide ester as well as a less reactive sulfonyl fluoride. The succinimide ester reacts rapidly with surface Lys residues “planting” the reagent at fixed locations on protein. The pendant aryl sulfonyl fluoride is then “cast” across a limited range of the protein surface, where it can react with multiple weakly nucleophilic amino acid sidechains in a proximity-enhanced sulfur-fluoride exchange (SuFEx) reaction. Using proteins of known structures, we demonstrated that the heterobifunctional agent formed cross-links between Lys residues and His, Ser, Thr, Tyr, and Lys sidechains. This geometric specificity contrasts with current bis-succinimide esters, which often generate nonspecific cross-links between lysines brought into proximity by rare thermal fluctuations. Thus, the current method can provide diverse and robust distance restraints to guide integrative modeling. This work provides a chemical cross-linker targeting unactivated Ser, Thr, His, and Tyr residues using sulfonyl fluorides. In addition, this methodology yielded a variety of cross-links when applied to the complex Escherichia coli cell lysate. Finally, in combination with genetically encoded chemical cross-linking, cross-linking using this reagent markedly increased the identification of weak and transient enzyme–substrate interactions in live cells. Proximity-dependent cross-linking will dramatically expand the scope and power of CXMS for defining the identities and structures of protein complexes.

scholarly journals The interaction between subunits in the tubulin dimer

1985 ◽  
Vol 230 (2) ◽  
pp. 551-556 ◽  
Author(s):  
L Serrano ◽  
J Avila
Keyword(s):  
Formic Acid ◽  
Limited Proteolysis ◽  
Trypsin Digestion ◽  
Cross Linking ◽  
Β Subunit ◽  
Α Subunit ◽  
Tubulin Dimer ◽  
Cross Linker ◽  
Chemical Cross Linking ◽  
Β Tubulin

Limited proteolysis and chemical cross-linking techniques have been used to study the interaction between α- and β-tubulin subunits. Trypsin digestion of tubulin dimer resulted in the cleavage of the α-subunit into two fragments, whereas chymotrypsin cleaved the β-subunit into two distinct fragments. All of these fragments have been mapped on the tubulin subunits by further proteolysis with formic acid. Cross-linking of trypsin- and chymotrypsin-cleaved subunits has been performed with two different cross-linker agents of different cross-linking distance. The addition of formaldehyde resulted in the cross-linking of the α-tubulin N-terminal fragment with β-tubulin C-terminal domain. The same result was obtained when methyl 4-mercaptobutyrimidate was used.

scholarly journals Some Properties of Chemical Cross-Linking Biohydrogel from Starch and Chitosan

2019 ◽  
Vol 2019 ◽  
pp. 1-6
Author(s):  
Titi Candra Sunarti ◽  
M. Irsan Febrian ◽  
Eka Ruriani ◽  
Indah Yuliasih
Keyword(s):  
Acid Concentration ◽  
Methacrylic Acid ◽  
Industrial Application ◽  
High Capacity ◽  
Monomer Conversion ◽  
Cross Linking ◽  
High Ability ◽  
Grafting Efficiency ◽  
Cross Linker ◽  
Chemical Cross Linking

Chemical cross-linking was developed to prepare starch and chitosan-based hydrogels. First, the precursor of starch was synthesized through the reaction of carboxymethylation with sodium monochloroacetate, and then chitosan was grafted by using methacrylic acid as cross-linker. In this research, sago and cassava starches were used and mixed with chitosan, and the effect of methacrylic acid concentration was investigated to determine the grafting parameters and hydrogel characteristics. Compared to native starch and carboxymethylated starch, hydrogels from both starches have high ability to swell and high capacity to absorb water and oil. The highest grafting yield, grafting efficiency, and monomer conversion were achieved by experiment using 0.550 g of methacrylic acid per g of CMS-chitosan mixture. These hydrogels have a good potency as biodegradable absorbents for pharmaceutical and industrial application.

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