title: Differential expression of P-type ATPases in intestinal epithelial cells: identification of putative new atp1a1 splice-variant.
abstract: P-type ATPases are membrane proteins that couple ATP hydrolysis with cation transport across the membrane. Ten different subtypes have been described. In mammalia, 15 genes of P-type ATPases from subtypes II-A, II-B and II-C, that transport low-atomic-weight cations (Ca(2+), Na(+), K(+) and H(+)), have been reported. They include reticulum and plasma-membrane Ca(2+)-ATPases, Na(+)/K(+)-ATPase and H(+)/K(+)-ATPases. Enterocytes and colonocytes show functional differences, which seem to be partially due to the differential expression of P-type ATPases. These enzymes have 9 structural motifs, being the phosphorylation (E) and the Mg(2+)ATP-binding (H) motifs the most preserved. These structural characteristics permitted developing a Multiplex-Nested-PCR (MN-PCR) for the simultaneous identification of different P-type ATPases. Thus, using MN-PCR, seven different cDNAs were cloned from enterocytes and colonocytes, including SERCA3, SERCA2, Na(+)/K(+)-ATPase alpha1-isoform, H(+)/K(+)-ATPase alpha2-isoform, PMCA1, PMCA4 and a cDNA-fragment that seems to be a new cassette-type splice-variant of the atp1a1 gen. PMCA4 in enterocytes and H(+)/K(+)-ATPase alpha2-isoform in colonocytes were differentially expressed. This cell-specific expression pattern is related with the distinctive enterocyte and colonocyte functions.
authors.affiliation: Lab. Fisiología Molecular, Centro de Biofísica y Bioquímica, Instituto Venezolano de Investigaciones Científicas, Apartado 20632, Caracas 1020-A, Venezuela. mrocaful@ivic.ve
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Rocafull
authors.forename: Luz E
authors.initials: LE
authors.lastname: Thomas
authors.forename: Girolamo J
authors.initials: GJ
authors.lastname: Barrera
authors.forename: Jesús R Del
authors.initials: JR
authors.lastname: Castillo
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D003106:Colon / Q000201:enzymology*; D020895:Enterocytes / Q000201:enzymology*; D006168:Guinea Pigs; D008969:Molecular Sequence Data; D000254:Sodium-Potassium-Exchanging ATPase / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000254:Sodium-Potassium-Exchanging ATPase