Phosphorylable proteins and alkaline phosphatase of brush border membranes from different parts of the rat small intestine.
1. The distribution along the small intestine of phosphorylable proteins from the brush border has been studied by gel electrophoresis. 2. Four proteins, with apparent Mr of 190, 160, 140 and 120 kDa were distributed unequally along the gut, which incorporated 32P from gamma 32P (ATP) to different degrees. 3. Alkaline phosphatase activity has been shown to follow the same distribution. 4. Under denaturing conditions 90, 85 and 65 kDa proteins were observed, whilst the proteins of 190, 160, 140 and 120 kDa had disappeared. 5. All these proteins, with the exception of the 190 kDa protein, had also been labelled with 32Pi. Furthermore, a difference in the phosphorylation of the 65 kDa and the 90-85 kDa proteins was observed. 6. The 65 kDa protein like commercial calf alkaline phosphatase had a ratio of phosphorylation from ATP to phosphorylation from Pi less than the 90 and 85 kDa proteins. 7. Mg2+ (2.5-10 mM) decreased phosphorylation of only the 65 kDa protein whilst beta-glycerophosphate inhibited phosphorylation of all forms of alkaline phosphatase. 8. Incorporation of gamma 32P (ATP) into the proteins was enhanced in the presence of 5 mM theophylline or EDTA. 9. The nature of the phosphorylation of these different proteins is discussed.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D003720:Densitometry; D004492:Edetic Acid / Q000494:pharmacology; D066298:In Vitro Techniques; D007421:Intestine, Small / Q000201:enzymology*; D008297:Male; D008871:Microvilli / Q000201:enzymology* / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism*; D051381:Rats
D010761:Phosphorus Radioisotopes; D011506:Proteins; D000255:Adenosine Triphosphate; D004492:Edetic Acid; D000469:Alkaline Phosphatase