Adipocyte differentiation of 3T3-L1 cells involves augmented expression of a 43 kDa plasma membrane fatty acid binding protein

A previously described 43-kDa plasma membrane fatty acid-binding protein (FABP(PM)) was not observed by immunohistochemical methods in proliferating 3T3-L1 fibroblasts. However, it was detectable in plasma membranes by the second day of confluent growth, prior to accumulation of visible lipid droplets, and was strongly expressed in 8-day differentiated adipocytes. These observations were confirmed by extraction of plasma membrane proteins and subsequent immunoblotting. Kinetics of initial [H-3]oleate uptake by both fibroblasts and adipocytes consisted of the sum of a saturable and a non-saturable component. During differentiation the saturable component increased progressively. V(max) increased from 3 to 25 to 110 pmol . s-1. mg cell protein-1 between the fibroblast, the 4-day, and 8 day adipocyte stages; K(m) was 24 nM in fibroblasts and approximately 55 nM in both 4- and 8-day differentiated adipocytes. By contrast, the rate constant for nonsaturable oleate influx decreased progressively from 0.026 to 0.010 ml. s-1 . mg protein-1 between the fibroblast and 8 day adipocyte stages. In 8-day adipocytes saturable oleate uptake was inhibited by up to 55% by antibodies against rat liver FABP(PM); these antibodies had no effect on uptake of 2-deoxyglucose or the medium chain fatty acid octanoate. They also had no effect on oleate uptake by fibroblasts. These studies support the hypothesis that FABP(PM) is a component of a saturable transport mechanism for long chain fatty acids.