Determination of specific radioactivity of amino acids in proteins directly on polyacrylamide gels: An application to l -leucine

1976 ◽  
Vol 72 (1-2) ◽  
pp. 577-585 ◽  
Author(s):  
A.F. Martin ◽  
G. Prior ◽  
R. Zak
1973 ◽  
Vol 51 (11) ◽  
pp. 2217-2222 ◽  
Author(s):  
R. B. van Huystee

The prime purpose of this proteolysis study was to direct attention to alternate means of measuring proteolytic activity other than the determination of free amino acids. The release of peptides from a macromolecular protein during incubation with either papain, pronase, or trypsin was determined by measuring the presence of 280-nm-absorbing molecules in the fractionation range of Sephadex G 25 eluant after incubation. The formation of larger proteinaceous constituents by proteolysis of arachin was analyzed by disc electrophoresis on polyacrylamide gels. Using these techniques it was noted that papain was the most efficient proteolytic agent for the degradation of arachin.


1985 ◽  
Vol 145 (1) ◽  
pp. 113-119 ◽  
Author(s):  
Laura L. Kilgore ◽  
Julie L. Rogers ◽  
Bruce W. Patterson ◽  
Nancy H. Miller ◽  
Waldo R. Fisher

1972 ◽  
Vol 128 (4) ◽  
pp. 879-899 ◽  
Author(s):  
Joseph Katz ◽  
P. A. Wals

1. Slices of mammary gland of lactating rats were incubated with glucose labelled uniformly with 14C and in positions 1, 2, 3 and 6, and with 3H in all six positions. Glucose carbon atoms are incorporated into CO2, fatty acids, lipid glycerol, the glucose and galactose moieties of lactose, lactate, soluble amino acids and proteins. C-3 of glucose appears in fatty acids. The incorporation of 3H into fatty acids is greatest from [3-3H]glucose. 3H from [5-3H]glucose appears, apart from in lactose, nearly all in water. 2. The specific radioactivity of the galactose moiety of lactose from [1-14C]- and [6-14C]-glucose was less, and that from [2-14C]- and [3-14C]-glucose more, than that of the glucose moiety. There was no randomization of carbon atoms in the glucose moiety, but it was extensive in galactose. 3. The pentose cycle was calculated from 14C yields in CO2 and fatty acids, and from the degradation of galactose from [2-14C]glucose. A method for the quantitative determination of the contribution of the pentose cycle, from incorporation into fatty acids from [3-14C]glucose, is derived. The rate of the reaction catalysed by hexose 6-phosphate isomerase was calculated from the randomization pattern in galactose. 4. Of the utilized glucose, 10–20% is converted into lactose, 20–30% is metabolized via the pentose cycle and the rest is metabolized via the Embden–Meyerhof pathway. About 10–15% of the triose phosphates and pyruvate is derived via the pentose cycle. 5. The pentose cycle is sufficient to provide 80–100% of the NADPH requirement for fatty acid synthesis. 6. The formation of reducing equivalents in the cytoplasm exceeds that required for reductive biosynthesis. About half of the cytoplasmic reducing equivalents are probably transferred into mitochondria. 7. In the Appendix a concise derivation of the randomization of C-1, C-2 and C-3 as a function of the pentose cycle is described.


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