Long-Chain-Fatty-Acid CoA Ligases: The Key to Fatty Acid Activation, Formation of Xenobiotic Acyl-CoA Thioesters and Lipophilic Xenobiotic Conjugates

Author(s):  
Kathleen Knights
1973 ◽  
Vol 131 (2) ◽  
pp. 199-209 ◽  
Author(s):  
J. Bar–Tana ◽  
G. Rose ◽  
R. Brandes ◽  
B. Shapiro

The mechanism of long-chain fatty acid activation catalysed by highly purified microsomal palmitoyl-CoA synthetase was investigated. The kinetics of the overall reaction were found to conform to the Bi Uni Uni Bi Ping Pong mechanism. 18O was transferred from [18O]palmitate to AMP and palmitoyl-CoA exclusively. The enzyme intermediate formed appeared to consist of enzyme-bound palmitate; this formation occurred only in the presence of ATP. However, the involvement of palmitoyl-AMP in the reaction catalysed by the purified enzyme has proved difficult to establish.


1972 ◽  
Vol 129 (5) ◽  
pp. 1101-1107 ◽  
Author(s):  
J. Bar–Tana ◽  
G. Rose ◽  
B. Shapiro

The partial and exchange reactions of long-chain fatty acid activation were determined by using purified microsomal long-chain fatty acyl-CoA synthetase (EC 6.2.1.3). No significant ATP formation from palmitoyl-AMP and PPi, nor palmitoyl-AMP-dependent CoA disappearance could be demonstrated. Similarly, no palmitate-dependent [32P2]PPi–ATP exchange was catalysed by the pure enzyme. The above partial and exchange reactions were, however, catalysed by the parent microsomal fraction at a rate similar to that of the overall reaction. The implications of these results are discussed.


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