Download e-book for iPad: Biological Oxidations by Th. Bücher (auth.), Professor Dr. Horst Sund, Professor Dr.

By Th. Bücher (auth.), Professor Dr. Horst Sund, Professor Dr. Volker Ullrich (eds.)

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Amino acid sequences used in study on thermal stability Protein Ferredoxin GAPDH Species References c. Tanaka et ale (1973) Tanaka et ale (1971) Azari et ale (1973) Rall et al. (1969) Tanaka et ale (1966) Tsunoda et ale (1968) C. P. C. C. M. ther-mosaaaha;rolytiaum tartarivoT'um elsdenii aaidi-uriai pasteurianum aerogenes T. aquatiaus B. stearothermophilus Yeast Pig Lobster LDH B. ~ I" ~ Fig. 4. Direction of preferred exchanges for producing a thermally stable enzyme. Arrows point from the mesophilic to the thermophilic protein.

7), as does the apo mouse LDH-X structure (Musick and Rossmann 1979a). The refined dogfish M4 apo LDH shows an intricate hy dro en bonding network around His 195 involving Asp 168, Asn 140, and S04- at the substrate binding site (Fig. 7). This network does not seem to exist in the ternary complex (White et al. 1976; Grau et al. 1981a) due to a conformational change of His 195, although these structures must still be refined to ascertain the accuracy of ' this assertion. Previous postulates on the mechanism have not taken into account such a "charge relay" system, Figure 8 shows a possible mechanism which includes the effect of Asp 168, the insertion of Arg 109 into the active center during catalysis, and the oil-water histidine proposal of Parker and Holbrook (1977) as modified by Grau et al.

I" ~ Fig. 4. Direction of preferred exchanges for producing a thermally stable enzyme. Arrows point from the mesophilic to the thermophilic protein. Numbers indicate the ranking of the significance for a given exchange '" Table 4. Benefit of exchange to stability Exchange Cold~Hot Gly~Ala Ser ~Ala Ser ~Thr Lys ~Arg Asp~Glu Ser ... Gly Lys .... Ala Val .... Ala Helical Region Sheet Region ++ ++ Internal Hydrophobicity External polarity ++ + + + ++ + ++ ++ + ++ + + ++ Asp~Asn Val~Ile + + + Internal Packing + ++ + The precise analysis of these data was given by Argos et al.

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