Nutrient transport in a bovine lens epithelial cell line

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Abstract
A bovine calf lens epithelial cell line (CLE‐1) that synthesizes crystallin has been established in culture and some of its transport properties have been characterized using both cells and membrane vesicles derived from them. The membrane vesicles fractionate with high recovery of plasma membrane markers, showing a 40‐fold purification of 5′‐AMPase and a 20‐fold decrease in the specific activity of the mitochondrial marker enzyme succinic dehydrogenase relative to a cell homogenate. Transport sites demonstrated higher specific activity than has been seen in vesicles from cell lines studied previously. The uptake of α‐amino isobutyric acid (AIB) (an alanine analog) by CLE‐1 cells is stimulated four‐ to fivefold by Na+ and exhibits a Km of 5.4 mM with a Vmax of 50 pmoles/min μg of cell protein. The uptake of leucine was not Na+ stimulatable. The uptake of AIB by the cells was reduced by 43% at confluence. Thus, the cell density dependent behavior of the uptake of the alanine amino acid family in CLE‐1 is similar to that of various fibroblast cells. The Na+ caused a threefold stimulation of AIB uptake in the membrane vesicles, while vesicular uptake of leucine was unaffected by Na+. The uptake of adenine, guanine, uridine, and guanosine was also tested in these vesicles. The substrates were rapidly accumulated, came to a steady state distribution within 1–2 minutes, and were recovered as the unaltered compounds after uptake.