Cysteine thiol

In addition, nonenzymatic acylation of cysteine thiols on proteins incubated in the presence of acyl-CoA has been described, although the biological importance of this process is still unclear. 

Figure 21-2. Fatty acid synthase multienzyme complex. The complex is a dimer of two identical polypeptide monomers, 1 and 2, each consisting of seven enzyme activities and the acyl carrier protein (ACP). (Cys— SH, cysteine thiol.) The— SH of the 4 -phosphopantetheine of one monomer is in close proximity to the— SH of the cysteine residue of the ketoacyl synthase of the other monomer, suggesting a "head-to-tail" arrangement of the two monomers. Though each monomer contains all the partial activities of the reaction sequence, the actual functional unit consists of one-half of one monomer interacting with the complementary half of the other. Thus, two acyl chains are produced simultaneously. The sequence of the enzymes in each monomer is based on Wakil.

Mordant dyes generally have the characteristics of acid dyes but with the ability in addition to form a stable complex with chromium. Most commonly, this takes the form of two hydroxy groups on either side of (ortho to) the azo group of a monoazo dye, as illustrated for the case of C. I. Mordant Black 1 (151). The dye is generally applied to the fibre as an acid dye and then treated with a source of chromium, commonly sodium or potassium dichromate. As a result of the process, the chromium(vi) is reduced by functional groups on the wool fibre, for example the cysteine thiol groups, and a chromium(m) complex of the dye is formed within the... 

Fig. 3 Chemistry of site-specific PEGylation developed by Brocchini et al. [5, 6]. After cleavage of the native disulfide bond between two cysteine thiols by reduction, the free cysteines are reacted with an a, (3-unsaturated PEG derivative to produce a PEG conjugate via a three-carbon bridge...

From a broader perspective, protein oxidation can result in covalent modification at many sites other than just at cysteine thiols. The earliest reports on protein oxidation date from the first decade of the twentieth century, but it took many more years to characterize these reactions and their products (Dakin, 1906). 

Figure 1.20 Cysteine and methionine are highly susceptible to oxidation reactions. Cysteine thiols can form disulfide linkages with other cysteine groups or be oxidized to cysteic acid. Methionine is oxidized very easily to the sulfoxide or sulfone products.

At the two extremes, lysine is observed as the amino acid most accessible on the surface of proteins while cysteine is the least exposed amino acid. The inaccessibility of cysteine probably stems from the fact that disulfides are typically buried within the polypeptide structure of proteins, whether they are intrachain or interchain in nature, and proteins rarely contain many reduced cysteine thiols. 

The protocol for using isobaric tags differs from that described previously for the ICAT or ECAT type reagents. In the following method, the proteins are denatured and the disulfides reduced and then alkylated to block them permanently. This eliminates disulfide re-association and also prevents the isobaric tags from forming thioester modification with cysteine thiols. Next, the proteins are digested with trypsin and then modified with an isobaric tag. Each sample is labeled with a different isobaric compound so that the samples can be differentiated upon MS/MS analysis. 

Another advantage to the use of a thiol additive is that the abundance of free thiol groups in the reaction environment will prevent the oxidation of the cysteine thiol at the N-terminal of the other peptide. Without added thiol transesterification catalysts, disulfide formation resulting in dimerization of the Cys-peptide would be a dominant side reaction in aqueous, oxygenated buffer conditions. 

Figure 22.16 SMPB-activated liposomes may be modified with peptide hapten molecules containing cysteine thiol groups. The resultant immunogen may be used for immunization purposes to generate an antibody...

Figure 28.21 The reactions of R u (11) pby 3 + are catalyzed by light at 452 nm that begins by forming an excited state intermediate. In the presence of persulfate, a sulfate radical is formed concomitant with the oxidative product Ru(III)bpy33+. This form of the chelate is able to catalyze the formation of a radical on a tyrosine phenolic ring that can react along with the sulfate radical either with a nucleophile, such as a cysteine thiol, or with another tyrosine side chain to form a covalent linkage. The result of this reaction cascade is to cause protein crosslinks to form when a sample containing these components is irradiated with light.

Scheme 13. Conjugate addition of cysteine thiol group to the maleiimido functionality for the synthesis of protein-peptide constructs...

In a comparison between various reactive dyes for wool and several commercially available colourless fibre-protecting agents, it was shown that reactive dyes in medium and full depths are significantly more effective. Reaction readily occurs between typical reactive dyes and the cysteine thiol groups (Scheme 7.47) released when the cystine disulphide bonds are hydrolysed under dyeing conditions. Such reactions inhibit thiol-disulphide... 

The nse of self-assembled monolayers of proteins by direct chemical attachment of the peptide to a modified snrface obviates some of the protein orientation difficnlties observed in the construction of LB mono-layers. An initial report of the construction of peptide SAMs on silanized qnartz was pnblished in early 1998 (185). Pilloud et al. have utilized the covalent ligation of protein cysteine thiols to thiol-terminated silanized... 

The reaction of an amino group with an aldehyde or ketone leads to an imine, which, as we have just seen with aldolase, provides a splendid example of how to bond a carbonyl substrate to an enzyme, and yet maintain its chemical reactivity in terms of enolate anion chemistry. Another type of covalent interaction is quite commonly encountered, and this exploits the thiol group of cysteine. Thiols are more acidic than... 

We have shown the cysteine thiol group as uncharged. The pAfa for this group in cysteine is about 10.3, and application of the Henderson-Hasselbach equation (see Section 4.9) indicates there will be negligible ionization at pH 7. Nevertheless, under the influence of a suitable basic group, e.g. arginine pATa 12.5, ionization to thiolate may be possible. In such an environment, thiolate may act as the nucleophile in the mechanism. 

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