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Isotopically-labeled substrates were incubated incubated with suspensions from r

ID: 967411 • Letter: I

Question

Isotopically-labeled substrates were incubated incubated with suspensions from respiring muscle tissue poisoned with malonate, blocking the citric acid cycle at reaction 6, succinate dehydrogenase. If the tricarboxylic acid cycle intermediates listed below are isolated, predict which carbon atoms of the isolated products would be expected to be labeled. Substrate is^14CH_3COCOOH with excess unlabeled oxaloacetate added. Products isolated are (1) isocitric acid, (2) a-ketoglutaric acid, (3) succinic acid. Substrate is^14CH_3COCOOH. Product is (1) isocitric acid, (2) a-ketoglutaric acid, (3) succinic acid. Substrate is H^14CO_3 Product is (1) isocitric acid, (2) a-ketoglutaric acid, (3) succinic acid, (4) malic acid. Substrate is 5-^14 C-fructose. Product is (l)isocitric acid, (2) succinic acid.

Explanation / Answer

         Aconitase

14CH3COCOOH + COOHCOCH2COOH                                                        COOH14CH2CHCOOCHOHCOOH

Pyruvic acid        Oxaloacetate                                     H2O                                  Isocitrate

H+

CO2   Isocitrate   DH

COOH14CH2CH2COCOOH

                                                                                                                                            - Keto glutarate

                                                -keto glutrate                                                                   fumarase

DH

COOH14CH2CH2COCOOH                           COOH14CH2CH2COOH                                   COOH14CH2CH2COOH

- Keto glutarate                                                                Succinic acid                                    Malic acid

Isotopically radio labelled isotopes incubated with suspensions from respiring muscle tissue poisoned with malonate block the citric acid cycle, radio labelled isotopes transfers their carbon atoms to 2 carbon atom. Fructose formed in gluconeogenesis cycle in the same way 2 carbon only.

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