Cysteine residues yeast

Affinity Labeling of Catalytic ATP Sites. Residues involved in ATP binding are potentially revealed by the use of affinity labels that are based on ATP s structure. Perhaps the most systematically studied of these compounds is 5 -fluorosulfonylbenzoyladenosine (5 -FSBA) (Figure 3a), which has been reported to label at least six kinases (32-A1). In the case of rabbit muscle pyruvate kinase such work has Indicated the presence of a tyrosine residue within the metal nucleotide binding site and an essential cysteine residue located at or near the free metal binding site (32). A similar reagent, 5 -FSBGuanosine, revealed the presence of two cysteine residues at the catalytic site of this same enzyme, both distinct residues from those modified by 5 -FSBA (33,34). With yeast pyruvate kinase both tyrosine and cysteine residues were modified by 5 -FSBA at the catalytic site ( ), and with porcine cAMP-dependent protein kinase a lysine residue was labeled at the active site (36). 

Fig. 11. Multiple sequence alignment of the copper chaperones for SODl using the CLUSTAL method (see text) (Higgins and Sharp, 1989). Sequence numbering corresponds to that of the yeast protein. Domain 1 cysteine residues believed to be important in uptake under copper-limiting conditions and domain 3 cysteine residues believed to be important in copper delivery to SOD 1 are boxed in black and labeled with asterisks. Residues in domain 2 that correspond to copper and zinc ligands in yeast and human SODl are boxed in black. Residues in domain 2 postulated to be involved in SODl target recognition (Trp-183 and Arg-217) are boxed in black and labeled with asterisks. The vertical arrows underneath domain 2 represent the positions of cysteine residues that make the disulhde bond in yeast and human SODl.

Fig. 14. X-ray crystal structure of full-length yeast CCS [pdb code Iqup (Lamb et al., 1999)]. (a) One monomer of yCCS is in light gray and the other is in dark gray. The cysteine residues of the MXCXXC motif in domain 1 are labeled and form a disulfide bond in each subunit. Amino acid side chains that are important in the formation of the positive patch at the dimer interface (Arg-188 and Arg-217) and the solvent-exposed Trp-183 residues of loop 6 at the center of this patch are shown in ball-and-stick representation. Domain 3 is not visible in the crystal structure (see text), (b) Stereo view of the image in (a) rotated 90° in the horizontal plane of the page and then 90° counterclockwise around an axis perpendicular to the page. The side chains that form the putative ySODl interaction surface are represented as ball-and-stick. The cysteine residues of the domain 1 MXCXXC motif are also represented in ball-and-stick.

Fig. 19. Multiple sequence alignment of Coxl7 proteins using the CLUSTAL method (Higgins and Sharp, 1989). Sequence numbering corresponds to that of the yeast protein. All cysteine residues in Saccharomyces cerevisiae Coxl7 and the seven invariant cysteine residues across all species are boxed in black. Cysteine residues that if mutated result in respiratory-dehcient yeast are labeled with asterisks.

The first evidence for this modification in mammalian cells was obtained when human Ha-Ras was transfected into rat fibroblasts and was found to be methylated at a C-terminal prenylated cysteine [38]. Based on the previous studies in yeast and jelly fungi, it was suggested this occurred by a third class of methyltransferase. Although Ras proteins have httle structural and sequence similarity to the fungi and yeast peptide substrates, they all have a unique characteristic a prenylated cysteine residue at the carboxyl terminus. The authors thus proposed that this motif is responsible for the recognition of this new class of methyltransf erase. 

NO-induced protein S-glutathionylation was proposed for the first time in 1988 by J.W. Park as a possible mechanism for the inactivation of yeast alcohol dehydrogenase by NO [32]. However, it took almost 10 years until the possibility that NO might be able to direct the incorporation of GSH to protein sulfhydryls was reconsidered. In 1997. it could be demonstrated that micromolar concentrations of GSNO inhibit aldose reductase through site-specific mixed disulphide formation at a conserved cysteine residue in its catalytic site... 

Free silver ions are the active components of antimicrobial silvers, and it has been shown that as little as one part per million of elemental silver in solution is an effective antimicrobial. Materials such as polymers, charcoal, and hydrocolloids when formulated with silver not only aid wound management and healing but also regulate its release into the wound environment and surrounding tissues. Silver ions kill micro-organisms by inhibiting cellular respiration and cellular function. " It is known that their mode of action is exerted by binding cysteine residues on the cell walls of yeasts such as C. albicans thereby... 

It is suggested that zinc is stored by metallothioneins in many tissues. These are low molecular weight proteins with a high incidence of cysteine residues. A zinc-binding protein, which is similar to metallothionein, has been isolated from yeasts and the cyanobacterium Anacystis nidulansf Zinc uptake in energy-starved Candida utilis requires protein synthesis. While it is possible that... 

SNAP-25 (synaptosomal-associated protein of 25kDa) is a 206 residue protein that lacks a classical transmembrane segment, but is bound to the cytosolic surface of the neuronal plasmalemma via pal-mitoylation of four cysteine residues located at the center of the molecule (Fig. 3) (Bennett and Scheller, 1994 Sudhof, 1995). SNAP-25 is required for axonal growth during development, and for nerve terminal plasticity in the mature nervous system (Osen-Sand et al., 1993). The tissue distribution of SNAP-25 is less well characterized than that of VAMP however, its presence in pancreatic cells (Jacobson et al., 1994 Sadoul et al., 1995) may indicate that it is also expressed outside the nervous system. A SNAP-25 related protein required for post-Golgi transport has been cloned from yeast (Brenwald efal., 1994). 

Inactivation of alcohol dehydrogenase from yeast with 14C-labeled [3-(3-bromoacetylpyridinio)-propyl]-adenosine pyrophosphate followed by oxidation showed the presence of 1-carboxymethyl histidine66. After inactivation of the enzyme with labeled [3-(4-bromoacetylpyridinio)-propyl]-adenosine pyrophosphate followed by oxidation, S-carboxymethyl cysteine was identified in the protein. In the case of glyceraldehyde-3-phosphate dehydrogenase, treatment with either coenzyme analogue leads to the modification of the cysteine residue. Treatment with [14C]nicotinamide-5-bromo-4-methylimidazole dinucleotide did not reveal any modified amino-acid-residues. The labeled nicotinamide residue split off during the recovery of the inactivated enzyme. Attempts to synthesize an inactivator labeled with a 14C-acetyl residue did not give satisfactory yields. If the enzyme-coenzyme derivative was treated with tritiated sodium boron hydride, tritium could be introduced (Fig. 22). Studies with... 

Bodmer et al. [11] have cloned, sequenced, and expressed active HGL in yeast The amino acid sequence obtained from the cDNA consists of 379 residues as compared to 449 residues constituting HPL. No amino acid sequence homology exists between HGL and HPL, except for short regions of six residues containing serine in a position analogous to that of the essential serine 152. The cDNA sequence of HGL shows the presence of three cysteine residues in comparison to PPL which contains 14 cysteine residues (6 disulfide bridges and 2 free sulfhydryl groups). 

A thioredoxin has also been partially purified from L. leichtnannii (39). Although this thioredoxin is similar in size (MW approximately 12,000) to the E. coli thioredoxin, it is not able to function as a substrate for the E. coli thioredoxin reductase. More recently thioredoxins have been purified to homogeneity from rat Novikoff ascites hepatoma (127) and from calf liver (45). The properties of these two thioredoxins are quite similar (MW 11,400 and 12,000 respectively). The amino acid compositions however, are different. For instance the Novikoff tumor thioredoxin contains six half-cysteine residues, whereas calf liver thioredoxin has only four. Both these thioredoxins have a tendency to aggregate in the oxidized form. This aggregation is probably due to mixed sulfide formation between the additional sulfhydryl groups. Like yeast thioredoxin II, calf liver thioredoxin contains only one tryptophan residue unfortunately the tryptophan content of Novikoff tumor thioredoxin was not determined. 

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