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G X Chen

Publications and source records attributed to G X Chen.

42 records · Page 3Linked to original sources
PubMed1981

Lipoic acid residues in a take-over mechanism for the pyruvate dehydrogenase multienzyme complex of Escherichia coli.

The pyruvate dehydrogenase complex of Escherichia coli contains two lipoic acid residues per dihydrolipoamide acetyltransferase chain, and these are known to engage in the part-reactions of the enzyme. The enzyme complex was treated with trypsin at pH 7.0, and a partly proteolysed complex was obtained that had lost almost 60% of its lipoic acid residues although it retained 80% of its pyruvate dehydrogenase-complex activity. When this complex was treated with N-ethylmaleimide in the presence of pyruvate and the absence of CoASH, the rate of modification of the remaining S-acetyldihydrolipoic acid residues was approximately equal to the accompanying rate of loss of enzymic activity. This is in contrast with the native pyruvate dehydrogenase complex, where under the same conditions modification proceeds appreciably faster than the loss of enzymic activity. The native pyruvate dehydrogenase complex was also treated with lipoamidase prepared from Streptococcus faecalis. The release of lipoic acid from the complex followed zero-order kinetics for most of the reaction, whereas the accompanying loss of pyruvate dehydrogenase-complex activity lagged substantially behind. These results eliminate a model for the enzyme mechanism in which specifically one of the two lipoic acid residues on each dihydrolipoamide acetyltransferase chain is essential for the reaction. They are consistent with a model in which the dihydrolipoamide acetyltransferase component contains more lipoic acid residues than are required to serve the pyruvate decarboxylase subunits under conditions of saturating substrates, enabling the function of an excised or inactivated lipoic acid residue to be taken over by another one. Unusual structural properties of the enzyme complex might permit this novel feature of the enzyme mechanism.

Amidohydrolases↗
PubMedDate not supplied

The oxidative folding of proteins by disulfide plus thiol does not correlate with redox potential.

The rate and yield of oxidative renaturation of reduced RNase A has been studied as a function of [-S-S-]/[-SH]. The principal conclusion of these studies is that rates and yields of oxidative renaturation are strongly dependent on the low mol. wt disulfide/thiol ratio. The relationships are complex and do not parallel the redox potential of the system. The present results are consistent with earlier findings on other proteins, and lead us to believe that the above conclusion is general. Kinetic studies of oxidative renaturation should recognize and account for the dependence of reaction rate and extent on the disulfide/thiol ratio. This ratio can change substantially over the course of a reaction, either due to stoichiometric transfer of disulfide to protein, and/or adventitious air oxidation of thiols. Failure to account for changes in the disulfide/thiol ratio may compromise the interpretation of such experiments.

Animals↗
PubMedDate not supplied

Bioartificial thyroid. The in vitro culture of microencapsulated rabbit thyroid tissue.

A new technique of encapsulating thyroid tissue with an artificial membrane for protection against immune attack is reported. The membrane was made of calcium alginate and a poly-L-lysin coating. The membrane prevents the diffusion of large molecules such as hemoglobin but freely permits the passage of smaller molecules such as T4 and culture medium 1640. Because larger molecules are barred, the contents of the microcapsules are safe from immune system rejection. In vitro cultured, microencapsulated rabbit thyroid tissue secreted T3 and T4, achieving a concentration of 8.68 +/- 2.93 nmol/L and 245.23 +/- 124.87 nmol/L, after 3 days; this level remained steady for 6 to 9 days of incubation. These microcapsules show promise as a treatment modality and as a possible solution to the problem of immuno-isolation.

Animals↗