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Chemical Structure| 115290-70-5 Chemical Structure| 115290-70-5

Structure of 115290-70-5

Chemical Structure| 115290-70-5

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Souza, Luiz HC ; Monteiro, Rayssa GF ; Guimarães, Wellinson G ; Gondim, Ana CS ; Sousa, Eduardo HS ; Diógenes, Izaura CN

Abstract: Oxidative stress on (Cys)-containing proteins has been associated with physiological disorders, as suggested for the human cofilin-1 (CFL-1) protein, in which the oxidized residues are likely implicated in the aggregation process of , leading to severe neuronal injuries. Considering the relevance of the oxidation state of , quantification of thiols may offer a guide for the development of effective therapies. This work presents, for the very first time, thiol quantification within CFL-1 in solution and on the surface following classic and adapted versions of Ellman’s assay. The 1:1 stoichiometric Ellman’sreaction occurs between 5,5′-dithio-bis(2-nitrobenzoic acid) (DTNB), and the free thiol of the residue, producing two 2-nitro-5-thiobenzoate (TNB2-) ions, one of which is released into the medium. While in solution, the thiol concentration was determined by the absorbance of the released TNB2-, on the surface, the mass of the attached TNB2- ion to the protein allowed the quantification by means of the multiparametric surface plasmon resonance (MP-SPR) technique. The SPR angle change after the interaction of with immobilized CFL-1 gave a surface coverage of 26.5 pmol cm−2 for the TNB2- ions (ΓTNB2-). The ratio of this value to the surface coverage of CFL-1, ΓCFL-1 = 6.5 ± 0.6 pmol cm−2 (also determined by MP-SPR), gave 4.1 as expected for this protein|https://www.ambeed.com/protein.html, i.e., CFL-1 contains four Cys residues in its native form (reduced state). A control experiment with adsorbed oxidized protein showed no SPR angle change, thus proving the reliability of adapting Ellman’s assay to the surface using the MP-SPR technique. The results presented in this work provide evidence of the heterogenization of Ellman’s assay, offering a novel perspective for studying thiol-containing species within proteins. This may be particularly useful to ensure further studies on drug-like molecules that can be carried out with validated oxidized or reduced CFL-1 or other analogous systems.

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Bartlett, Ryan ;

Abstract: Synthesis of small-molecule conjugates capable of selective attachment to a peptide/protein backbone paves way for new therapeutic applications of protein modification. Sufficient inhibition or installation of function requires extensive design so the compound may identify and attach to the intended site. The utility of such an approach has prompted a number of applicable strategies and has rendered the development of new modification scaffolds less necessary. On the other hand, covalent modification to the biologics with ability to remove the conjugate via stimulus is underdeveloped, amplifying a current need to investigate the reversible potential for conjugation and deconjugation techniques in regard to molecular scaffolds. The investigation reported therein involves a small molecule design capable of conjugation and facile deconjugation from peptide/protein structure. Taking advantage of special reactivity between aldehyde and thiol, we utilize a number of readily available mercapto-benzaldehyde derivatives that have been synthesized by simple procedure and characterized by NMR. These compounds chemoselectively bind to backbone amino by institution of distinctive [3.3.1] and [2.2.1] bicyclic scaffolds. This protocol enables amino modification in a multi-component fashion in aqueous buffer and mild conditions, characterized by combination of HPLC and LC-MS methods. The various bicyclic modification derivatives were investigated for progression of the reversed reaction under mild condition, utilizing a commercially available amine reagent. In restoring the original protein motif, a full investigation of a heterocyclic derivative for reversible attachment to several linear peptides was subsequently performed. Upon completion, the effectiveness of this approach was demonstrated in the modification and quantifiable regeneration of insulin protein.

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Product Details of [ 115290-70-5 ]

CAS No. :115290-70-5
Formula : C9H15O6P
M.W : 250.19
SMILES Code : O=C(CCP(CCC(O)=O)CCC(O)=O)O
MDL No. :MFCD12068446

Safety of [ 115290-70-5 ]

GHS Pictogram:
Signal Word:Danger
Hazard Statements:H314
Precautionary Statements:P301+P330+P331-P303+P361+P353-P363-P304+P340-P310-P321-P260-P264-P280-P305+P351+P338-P405-P501
Class:8
UN#:3265
Packing Group:
 

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