Disposition of the carboxy-terminus tail of rabbit lactase-phlorizin hydrolase elucidated by phosphorylation with protein kinase A in vitro and in tissue culture.
The intracellular disposition of the carboxy-terminus tail of rabbit lactase-phlorizin hydrolase (LPH) is demonstrated, using a specific phosphorylation of Ser1916 by protein kinase A (PKA). This phosphorylation is shown to occur not only in vitro (with pure LPH and pure catalytic subunit of PKA), but also in an organ culture of the small intestine. Cholera toxin, which is known to act in vivo on the membranes of the small intestine, with severe clinical consequences, and to elevate the intracellular cyclic AMP of enterocytes, is shown to enhance significantly the phosphorylation of LPH in intact cells grown as an organ culture. These findings establish the cytosolic orientation of the carboxy-terminus tail of LPH in situ, and raise the possibility that the tail itself and its phosphorylation by PKA may have a physiological or physiopathological significance.
Keller
P
P
Department of Biochemistry, Swiss Federal Institute of Technology, ETH Zentrum, Zurich.
Semenza
G
G
Shaltiel
S
S
D000595:Amino Acid Sequence; D000818:Animals; D046508:Culture Techniques; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D006801:Humans; D010694:Lactase-Phlorizin Hydrolase / Q000737:chemistry* / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011817:Rabbits; D017386:Sequence Homology, Amino Acid
D017868:Cyclic AMP-Dependent Protein Kinases; D010694:Lactase-Phlorizin Hydrolase