Molecular cloning and sequencing of the cDNA coding for a calcium‐binding protein regucalcin from rat liver
- 2 August 1993
- journal article
- Published by Wiley in FEBS Letters
- Vol. 327 (3), 251-255
- https://doi.org/10.1016/0014-5793(93)80998-a
Abstract
The cDNA of a Ca2+‐binding protein regucalcin was cloned from a rat liver cDNA library which was constructed in λZAPII by immunoscreening. Positive clones were obtained from which spanned the region of interest, and they gave a sequence of 1.7 kb by sequencing with the dideoxynucleotide method. Analysis of the sequence of the cloned cDNA showed that the cDNA encoded the complete amino acid sequence of the regucalcin molecule. Regucalcin was composed of 299 amino acid residues and its molecular weight was estimated to be 33,388 Da. The hydropathy profile of regucalcin showed a highly hydrophilic character. The nucleotide and amino acid sequences of regucalcin did not have statistically significant homology, as compared with the registered sequences which are found in the EMBL and GenBank databases containing several other Ca2+‐binding proteins (calmodulin, calbindin‐D28k and S‐100β). The regucalcin molecule did not contain the EF‐hand motif as a Ca2+‐binding domain. The present study demonstrates that regucalcin is a unique Ca2+‐binding protein in the liver of rats.Keywords
This publication has 22 references indexed in Scilit:
- Inhibitory effect of calcium-binding protein regucalcin on Ca2+/calmodulin-dependent cyclic nucleotide phosphodiesterase activity in rat liver cytosolMolecular and Cellular Biochemistry, 1991
- Inhibitory effect of calcium‐binding protein regucalcin on Ca2+‐activated DNA fragmentation in rat liver nucleiFEBS Letters, 1991
- Calcium-Binding Proteins: Past, Present and FuturePublished by Springer Science and Business Media LLC ,1991
- λ ZAP: a bacteriophage λ expression vector within vivoexcision propertiesNucleic Acids Research, 1988
- Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extractionAnalytical Biochemistry, 1987
- Dideoxy sequencing method using denatured plasmid templatesAnalytical Biochemistry, 1986
- A simple and very efficient method for generating cDNA librariesGene, 1983
- Properties of calcium-binding protein isolated from the soluble fraction of normal rat liver.CHEMICAL & PHARMACEUTICAL BULLETIN, 1981
- Purification of calcium binding substance from soluble fraction of normal rat liver.CHEMICAL & PHARMACEUTICAL BULLETIN, 1978
- DNA sequencing with chain-terminating inhibitorsProceedings of the National Academy of Sciences of the United States of America, 1977