Cycle of VIP in the human transformed colonic epithelial cells (HT-29) in culture

The kinetics of VIP processing in different compartments (medium, plasma membrane and intracellular) by HT-29 cells was studied using direct biochemical methods and a homogeneous monoiodinated VIP. The compartmental radioactivity was characterized by HPLC fractionation and specific receptor binding. VIP once bound to the cell surface remains intact and is rapidly and maximally internalized (less than 10 min) at 37 degrees C. Then two different processes occur: (1) release of degradation products 125I and monoiodinated tyrosine in the medium; (2) VIP remains intact in the cells representing 67.2 +/- 4.7% of total radioactivity up to 90 min. The overall processing of VIP is time- and temperature-dependent and maximal internalization of VIP with minimal medium release is observed at 20 degrees C. Our results demonstrate a receptor mediated internalization of VIP and that at least two intracellular pathways may exist in the cycle of VIP. One is associated with a complete degradation of VIP detected in the extracellular medium and is optimal at 37 degrees C. The other results in the presence of intact intracellular VIP and is optimal at 20 degrees C.