The uptake of swainsonine, a specific inhibitor of α-D-mannosidase, into normal human fibroblasts in culture

✍ Scribed by Kokila Chotai; Christine Jennings; Bryan Winchester; Peter Dorling

• Publisher: John Wiley and Sons
• Year: 1983
• Tongue: English
• Weight: 653 KB
• Volume: 21
• Category: Article
• ISSN: 0730-2312
• DOI: 10.1002/jcb.240210202

✦ Synopsis

Swainsonine, an indolizidine alkaloid, found in plants of the genus Swainsona, has been shown to be a strong inhibitor in vitro of the a-D-mannosidase activity in normal human fibroblasts. Therefore, inhibition of a-D-mannosidase activity in extracts of harvested cells grown with swainsonine in the medium has been used to follow the association of the alkaloid with normal human fibroblasts in culture. Swainsonine that could not be removed by extensive washing became associated with the cells within 1 min, and it is concluded that the alkaloid is internalized rapidly by the cells. The amount of swainsonine taken up into the cells depended on the length of time in contact and the concentration of swainsonine in the medium, but at 37°C a plateau of internalized swainsonine occurred after 2 hr with extracellular concentrations of swainsonine of 100 p M or greater.

At lower concentrations of swainsonine the rate of uptake was found to be temperature-dependent, increasing greatly at 20°C. The rapidity and temperature sensitivity of the uptake, together with the observation that mannose or mannose-6-phosphate did not prevent the association, suggest that swainsonine enters the cells by permeation rather than by endocytosis. When swainsonine is withdrawn from the culture medium, there is a decrease with time of cell-associated swainsonine. The kinetics of uptake and release of swainsonine and its slightly basic nature make it likely that swainsonine is concentrated initially in the lysosomes. This rapid, but reversible, concentration of swainsonine in lysosomes would be consistent with the observed effects of the toxin in vivo.