Plasma-membrane calcium-pump isoforms in human and rat liver.
Plasma-membrane Ca(2+)-pumping ATPases (PMCAs) extrude Ca2+ from the cytoplasm of all cells. Some previous studies of ATP-dependent Ca2+ transport by liver membranes suggested there exist specific properties of the hepatic PMCA, including regulation by hormones which affect calcium signalling. Multiple PMCA isoforms are now known to result from expression of four different genes (known as PMCA 1-4) and alternative RNA splicing at three possible sites (A, B and C). We investigated which isoforms are expressed in adult human and rat liver RNA using reverse-transcription polymerase chain reaction with mixed primers designed to amplify parts of all the known PMCA transcripts. In human liver, products were identified by sequencing from PMCA1, PMCA2 and PMCA4, but not from PMCA3 or from any new gene. In rat liver, by contrast, only PMCA1 and PMCA2 were detectable, although we confirmed that the primers were able to amplify from rat lung a new sequence which is part of rat PMCA4. Of the alternatively spliced variants, at site A in the PMCA2 sequences, all the exons were included in both adult and fetal human liver. In human liver, the exon at site B was excluded in some products from PMCA1 and PMCA4, and at site C, only PMCA1b and one form of PMCA4 were found. Blots of human liver RNA showed PMCA1 and PMCA4 were abundantly expressed, unlike PMCA2. On blots of rat liver RNA, PMCA1 was more abundant than PMCA2, and purified rat parenchymal cell RNA gave similar findings. In summary, no new hepatic PMCA isoforms have been demonstrated, but differences between the predominant human and rat isoforms may have consequences for Ca2+ signalling or the response to liver cell injury.
Howard
A
A
Department of Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London, U.K.
Barley
N F
NF
Legon
S
S
Walters
J R
JR
D000328:Adult; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D000252:Calcium-Transporting ATPases / Q000096:biosynthesis* / Q000235:genetics; D002462:Cell Membrane / Q000201:enzymology; D017931:DNA Primers; D015342:DNA Probes; D005333:Fetus; D015870:Gene Expression; D014644:Genetic Variation; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D051381:Rats
D017931:DNA Primers; D015342:DNA Probes; D007527:Isoenzymes; D000252:Calcium-Transporting ATPases