The internalization mechanisms and trafficking of the pea albumin in Caco-2 cells

Abstract

Pea albumin (PA) can reach the intestine in the active form because it is highly resistant to gastric acid and proteolytic enzymes after their oral intake, which can supply various bioactivities. However, there is no detailed knowledge of the intestinal cell uptake about PA. The aim of this work was to study the internalization mechanism and intracellular trafficking route of PA. The uptake of PA-cyanine 5.5 NHS ester (Cy5.5) was a time-dependent and concentration-dependent process in Caco-2 cells. Endocytosis inhibitors or small interfering RNA (siRNA) techniques revealed that the internalization of PA-Cy5.5 was energy-dependent and mediated by caveolin-mediated endocytosis. Furthermore, we observed colocalization of PA-Cy5.5 and its subcellular localization in Caco-2 cells by using confocal laser scanning microscopy, which revealed that the intracellular trafficking process of PA-Cy5.5 was related to endoplasmic reticulum, Golgi, and lysosome. Interestingly, PA can alleviate lipopolysaccharide -induced ER stress, which may be the main reason why pea albumin is anti-inflammatory. Overall, our findings suggest caveolin may be critical for PA uptake in enterocytes and could contribute to explore the bioactivities mechanism of pea albumin in body.