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5 . Other Reagents Keech and Farrant (95) have reported that l-fluor0-2~4-dinitrobenzene reacts with the c-amino group of a single lysyl residue in pyruvate carboxylase purified from sheep kidney cortex causing inactivation of this enzyme. Since only acetyl-CoA among the reaction components protects against this inactivation, the lysyl residue attacked appears to be implicated either in the binding of acetyl-CoA or in mediation of the effect(s) of this activator on the catalytic site ( 9 5 ) . Similar effects are not however observed when the r-amino groups of pyruvate carboxylase from avian liver are modified using the specific reagent, 2-rnethoxy-5nitrotropone (46, 9 6 ) .

Marshall, R. 1,. Metzenberg, and P. P. Cohen, JBC 236, 2229 (1961). 101. J. J. Cazzulo, T. K. Sundaram, and H. L. Kornberg, Proc. Roy. Soc. B176, 1 (1970). f . Require& Requiredd 3 2 Acetyl-CoA Acetyl-CoA No No (1,108) Requiredd Stimulatorye None None 3 1 - Acetyl-CoA Palmityl-CoA - Yes YeS YeS No (1011 (6, 77, 98) (37) (10,23) (5,80,103,104) Obtained from Eq. (6) (105): T I * On the basis of K A , the concentration of acyl-CoA required to achieve 50% maximal activation. Initial rate of CO, fixation is increased more than 1000-fold in the presence of a saturating concentration of acetyl-CoA (102).

E. C. Wacker, in “The Proteins” (H. , Vol. 5, p. 25. Academic Press, New York, 1970. 64. M. Cohn. Biocliernlstry 2, 623 (1963). 65. A. S. Mildvan, J. S. Lcigh, and M. Cohn, Biochemistry 6, 1805 (1967). 66. A . S. Mildvan and M. Cohn, Advan. Enzymol. 33, 1 (1970). 67. M. C. Scrutton and A. S. Mildvan, in “CO,: Chemical, Biochemical and Physiological Aspects” (J. T. ), NASA-SP188, p. 207. U. S. Govt. Printing Office, Washington, D. , 1970. 12 M . C. SCRUTTON AND M. R. YOUNG a. Effects on the Nuclear Magnetic Relaxation Rates of Water Protons.

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