Solubilization of C18-CoA and C20-CoA elongases from Allium porrum L. epidermal cell microsomes

• Published in: FEBS letters 1985, Vol.187 (2), p.314-320
• Main Authors: Lessire, R, Bessoule, J.J, Cassagne, C
• Format: Article
• Language: English
• Subjects: acyl coenzyme a; acyl-CoA; Allium porrum; Analytical, structural and metabolic biochemistry; Biological and medical sciences; coenzyme A; enzyme activity; enzymes; Enzymes and enzyme inhibitors; esters; fatty acid esters; Fundamental and applied biological sciences. Psychology; isolation; Ligases; lipogenesis; long chain fatty acids; microsomes; nonionic surfactants
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(85)81267-9

The effects of n-octyl-beta-D-glucopyranoside, Triton X-100 and deoxycholate on acyl-CoA elongation by Allium porrum L. epidermal cell microsomes showed that the Triton X-100 specifically stimulated the synthesis of C22-C26 acids using C18-CoA as primer, whereas the fatty acid elongation products of C20-CoA remained essentially unchanged. n-Octyl-beta-D-glucopyranoside increased the C20 and C22 fatty acid syntheses to the same extent and deoxycholate inhibited C18-CoA and C20-CoA elongation. The presence of two different elongation systems, as suggested by these results, has been demonstrated. After solubilization by Triton X-100, the C18-CoA and C20-CoA elongases were separated by sucrose density centrifugation. The fractions corresponding to sucrose concentrations of 0.51 and 0.62 M presented the maximal activities for C18-CoA and C20-CoA elongases, respectively. In addition, by gel filtration on a Sephacryl S-300 column, the C20-CoA and the C18-CoA elongases have estimated apparent molecular masses under detergent conditions of 600 and 350 kDa, respectively.