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By Efraim Racker, Anthony San Pietro

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This mode of attachment anchors the acyl group firmly to the enzyme and prevents phosphate or arsenate from reacting with the enzyme in the usual manner. This concept requires the effectiveness of this inhibitor to be dependent on the intactness of the phosphate transfer site. In the next lecture we shall see that this is the case. LECTURE 3 SUBSTRATE LEVEL OXIDATIVE PHOSPHORYLATION (B) A scientist living at Staines Is searching with infinite pains For a new type of sound Which he hopes, when it's found, Will travel much faster than planes.

It also explains the curious discrepancies in the Km values for D P N recorded in the litera24 J. Harting, in "The Mechanism of Enzyme Action" (W. D. McElroy and B. ), p. 536. Johns Hopkins Press, Baltimore, 1954. 3. Substrate Level Oxidative Phosphorylation 34 ture. As shown in Fig. 7 mM). If D P N H is removed by pyruvate and lactate dehydrogenase, the oxidation of glyceraldehyde 3-phosphate is rapid and zero order even at the low DPN concentrations. 2 C 0 1 1 2 I 3 1 4 1 5 I 6 I 7 I MINUTES F I G .

21 Phosphoglyceryl-enzyme was prepared in the presence of a regenerating system for 1,3-diphosphogly cerate and was passed through a Sephadex column. The emerging protein was analyzed and found to have lost more than 75% of its DPN content. Control experiments with untreated enzyme, or with enzyme treated with the same mixture of reagents except for ATP, which was omitted, revealed no loss of DPN. A lowered affinity to D P N after acylation of the enzyme is also apparent from the requirement for high DPN concentration for optimal rates of arsenolysis of 1,3-diphosphoglycerate.

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