Yeast enolase

Lebioda, L., Stec, B., Brewer, J.M. The structure of yeast enolase at 2.5 A resolution. An 8-fold p + a barrel with a novel ppoa(pa)6 topology. /. Biol. Chem. 

Yeast alcohol dehydrogenase, 5. 1009 cobalt-containing, 5, 1013 manganese-containing, S, 1014 Yeast enolase activation... 

Larsen, T.M., Wedeking, J.E., Rayment, I. and Reed, G.H. (1996) A carboxylate oxygen of the substrate bridges the magnesium ions at the active site of enolase structure of the yeast enzyme complexed with the equilibrium mixture of 2-phosphoglycerate and phosphoenolpyruvate at 1.8 A resolution, Biochemistry, 30, 4349-4358. 

The Truhlar group has reported an interesting theoretical study of H/D kinetic isotope effects for conversion of 2 phospho-D-glycerate to phosophoenolpyruvate catalyzed by the yeast enolase enzyme. The proton transfer step (first reaction step in Fig. 11.10) is the rate limiting step and was chosen for theoretical study. The KIE for proton/deuteron transfer is kn/kD = 3.3 at 300 K. 

Nussbaum, a. K., Dick, T. P., Keilholz, W., Schiele, M., Stevanovic, S., Dietz, K., Heinemeyee, W., Geoll, M., Wole, D. H., Hubee, R., et al. Cleavage motifs of the yeast 20S proteasome beta subunits deduced from digests of enolase 1. Proc. Natl. Acad. Sci. USA 1998, 95, 12504-12509. 

FIGURE 2. Left deprotonation of pVIg(OH2)6] lowers the coordination number of magnesium. Reprinted with permission from Reference 14. Copyright 2005 American Chemical Society. Right pentacoordinate geometry of Mg + in the solid-state structure of the Mg + —F —Pi complex of yeast enolase... 

Since then, a considerable amount of structural and mechanistic information has been collected and yeast enolase is probably the best understood sequential enzyme to date. It is a homodimer and requires two Mg + ions per active site for catalytic activity under physiological conditions, although magnesium can be replaced with a variety of divalent metal ions in vitro. During a catalytic turnover, the metal ions bind to the active site in a kinetically ordered, sequential manner with differential binding affinities. The mode of action of yeast enolase is illustrated in Figure 26 and is unusually well understood since several solid-state structures for each intermediate identified with kinetic methods have been determined. 

FIGURE 26. Above the kinetically ordered sequential mode of action of yeast enolase. Reprinted with permission from Reference 250. Copyright 2001 with permission of the American Chemical Society. Below conformational changes in the chemical step... 

PLATE 5 Loop movement in the active site of yeast enolase. Lfpper left closed conformation (PDB 2AL1) superimposed upon the open conformation (PDB 1P43). Upper right view from the back. Lower left A quantum chemical soccer ball model for yeast enolase illustrated on the enol-intermediate and calculated at the TPSS(MARI-J COSMO)/SV(P) level of theory. Lower right view from the back... 

Case Study Catalytic Mechanism of Yeast Enolase... 

CASE STUDY CATALYTIC MECHANISM OF YEAST ENOLASE... 

Yeast enolase (Mr 93,316) is a dimer with 436 amino acid residues per subunit. The enolase reaction illustrates one type of metal ion catalysis and provides an additional example of general acid-base catalysis and transition-state stabilization. The reaction occurs in two steps (Fig. 6-23a). First, Lys345 acts as a general base catalyst,... 

Figure 13-2 View of the active site of yeast enolase containing a bound molecule of 2-phospho-D-glycerate. The catalytic magnesium ion is at the left but the "conformational" metal is not visible here. The imidazole group of His 159 serves as the catalytic base and the -NH3+ of Lys 396 or Lys 34573b as the catalytic acid. From Vinarov and Nowak.69...

Nittner-Marszalska, M., Wojcicka-Kustrzeba, I., Bogacka, E., Patkowski, J., and Dobek, R. 2001. Skin prick test response to enzyme enolase of the baker s yeast (Saccharomyces cerevisiae) in diagnosis of respiratory allergy. Med Sci. Monit 7(1) 121-124. 

FIGURE 18 Countercurrent distribution of enolase from baker s yeast ( ) experimental data (O) theoretical (—) sum of theoretical curves. (From G. Blomquist and S. Wold, Numerical resolution of CCD [counter current distribution] curves. Acta Chem. Scand. B28, 56-60, 1974.)... 

Second derivative spectroscopy is a means to extract information from the spectrum. In contrast to the relatively featureless UV spectra, the second derivatives of UV spectra are characterized by 2 sharp peaks and troughs [6,7]. Figure 1 shows the ultraviolet spectra of yeast enolase at 0.28 and 200 MPa and Figure 2 shows the second derivative of those spectra. 

Holland MJ, Holland JP, Thill GP, Jackson KA The primary structures of two yeast enolase genes. Homology between the 5 noncoding flanking regions of yeast enolase and glyceraldehyde-3-... 

Baldo BA, Baker RS Inhalant allergies to fungi Reactions to bakers yeast Saccharvmyces cerevisiae) and identification of bakers yeast enolase as an important allergen. Int Arch Allergy Appl Immunol 1988 86 201-208. 

Lebioda L, Stec B, Brewer JM The structure of yeast enolase at 2.25-A resolution. An 8-fold beta + alpha-barrel with a novel beta beta alpha alpha (beta alpha)6 topology. J Biol Chem 1989 264 3685-3693. 

Stec B, Lebioda L Refined structure of yeast apo-enolase at 2.25 A resolution. J Mol Biol 1990 211 235-248. 

Westhead EW Enolase from yeast and rabbit muscle. Methods Enzymol 1966 9 670-679. 

F) Active site of lobster enolase with a bound phosphoglycolate inhibitor and those side chains involved in specific interactions (IPDZ, Plate XXXI), showing an El structure shghtly different from that of the yeast enzyme bound with the same inhibitor in (C), wherein the inhibitor is bound to the metal... 

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