Molecular cloning of a plasma membrane calcium pump from human osteoblasts.
The osteoblast plays a critical role in bone formation, bone remodelling, bone matrix formation, and matrix calcification. To better understand the process of osteoblast-controlled bone formation, we determined the structure and isoform types of the plasma membrane calcium pump from normal human osteoblasts. A complementary DNA library from normal human osteoblasts was screened for plasma membrane calcium pump clones. Sequencing and analysis of cDNA clones revealed the presence of a 3986 base pair cDNA that encoded a 1220 amino acid protein that was similar to the human plasma membrane calcium pump isoform 1. Polyadenylated RNA from human osteoblast cells contains bands of RNA approximately 5050 and 6750 bases long. Reverse transcription of polyadenylated RNA from human osteoblasts followed by amplification of the RNA-DNA duplex with calcium pump isoform-specific primers revealed the presence of isoforms 1 and 2 of the calcium pump. Isoform 4 was not detected. We conclude that normal adult human osteoblasts contain a plasma membrane calcium pump that is similar to the human plasma membrane calcium pump isoform 1. It is likely that this pump plays an important role in the cell biology of the human osteoblast.
Nephrology Research Unit, Mayo Clinic and Foundation, Rochester, Minnesota.
R
R
Kumar
J D
JD
Haugen
J T
JT
Penniston
D000328:Adult; D000595:Amino Acid Sequence; D001483:Base Sequence; D000252:Calcium-Transporting ATPases / Q000235:genetics*; D002462:Cell Membrane / Q000737:chemistry; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D010006:Osteoblasts / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D004247:DNA; D000252:Calcium-Transporting ATPases