Primary structure and functional expression of the Na/Ca,K‐exchanger from bovine rod photoreceptors.

Complementary DNA encoding the Na/Ca,K‐exchanger was isolated from bovine retina cDNA libraries. The complete full‐length cDNA is approximately 4 kb long and contains an open reading frame of 3597 bp. The deduced amino acid sequence corresponds to a protein of 1199 amino acids with a calculated molecular weight of approximately 130 kDa. Hydrophobicity analysis revealed the presence of two alternating sets of hydrophobic and hydrophilic domains. There also exists a hydrophobic region at the N‐terminus which may be part of a cleavable signal peptide. The protein shares limited sequence homology with the Na/Ca‐exchanger from cardiac sarcolemma. Northern blot analysis indicates that the approximately 6 kb transcript is highly specific for retinal tissue. Insect cells infected with recombinant baculovirus bearing the full‐length cDNA express a functional Na/Ca,K‐exchanger with an apparent relative molecular weight of approximately 210 kDa, as determined by Western blotting.

[1]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[2]  H. Michel,et al.  Expression and cell membrane localization of rat M3 muscarinic acetylcholine receptor produced in Sf 9 insect cells using the baculovirus system , 1991, FEBS letters.

[3]  H. Rahamimoff,et al.  The modulation of rat brain Na(+)-Ca2+ exchange by K+. , 1991, The Journal of biological chemistry.

[4]  G. Wolbring,et al.  The sodium-calcium exchanger of bovine rod photoreceptors: K(+)-dependence of the purified and reconstituted protein. , 1991, Biochimica et biophysica acta.

[5]  D. Nicoll,et al.  Molecular cloning and functional expression of the cardiac sarcolemmal Na(+)-Ca2+ exchanger , 1990, Science.

[6]  W Haase,et al.  Immunological characterization and localization of the Na+/Ca2(+)- exchanger in bovine retina , 1990, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[7]  Ylva Gavel,et al.  Sequence differences between glycosylated and non-glycosylated Asn-X-Thr/Ser acceptor sites: implications for protein engineering , 1990, Protein engineering.

[8]  R. Molday,et al.  Identification of the sodium-calcium exchanger as the major ricin-binding glycoprotein of bovine rod outer segments and its localization to the plasma membrane. , 1990, Biochemistry.

[9]  L. Tsui,et al.  Erratum: Identification of the Cystic Fibrosis Gene: Cloning and Characterization of Complementary DNA , 1989, Science.

[10]  P. Schnetkamp,et al.  Na+-Ca2+ exchange in bovine rod outer segments requires and transports K+. , 1989, The American journal of physiology.

[11]  C. Slaughter,et al.  Adenylyl cyclase amino acid sequence: possible channel- or transporter-like structure. , 1989, Science.

[12]  H. Takeshima,et al.  Primary structure and expression from complementary DNA of skeletal muscle ryanodine receptor , 1989, Nature.

[13]  L. Lagnado,et al.  Extrusion of calcium from rod outer segments is driven by both sodium and potassium gradients , 1989, Nature.

[14]  G. Del Sal,et al.  A one-tube plasmid DNA mini-preparation suitable for sequencing. , 1988, Nucleic acids research.

[15]  F. Lottspeich,et al.  A new siliconized-glass fiber as support for protein-chemical analysis of electroblotted proteins. , 1988, European journal of biochemistry.

[16]  U. Kaupp,et al.  Solubilization, purification, and reconstitution of the sodium-calcium exchanger from bovine retinal rod outer segments. , 1988, The Journal of biological chemistry.

[17]  L. Lagnado,et al.  Ion transport by the Na-Ca exchange in isolated rod outer segments. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[18]  K. Mullis,et al.  Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. , 1988, Science.

[19]  M. Kozak.  An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. , 1987, Nucleic acids research.

[20]  A. D. McLachlan,et al.  Profile analysis: detection of distantly related proteins. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[21]  U. Kaupp,et al.  Solubilization and functional reconstitution of the cGMP-dependent cation channel from bovine rod outer segments. , 1986, The Journal of biological chemistry.

[22]  D. Housman,et al.  Mammalian multidrug resistance gene: Complete cDNA sequence indicates strong homology to bacterial transport proteins , 1986, Cell.

[23]  G. Heijne.  A new method for predicting signal sequence cleavage sites. , 1986 .

[24]  P. Schnetkamp.  Sodium‐calcium exchange in the outer segments of bovine rod photoreceptors. , 1986, The Journal of physiology.

[25]  E. E. Fesenko,et al.  Induction by cyclic GMP of cationic conductance in plasma membrane of retinal rod outer segment , 1985, Nature.

[26]  K. Yau,et al.  Electrogenic Na–Ca exchange in retinal rod outer segment , 1984, Nature.

[27]  S. Henikoff.  Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. , 1984, Gene.

[28]  K. Yau,et al.  Cation selectivity of light-sensitive conductance in retinal rods , 1984, Nature.

[29]  M. Kozak.  Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. , 1984, Nucleic acids research.

[30]  J. Devereux,et al.  A comprehensive set of sequence analysis programs for the VAX , 1984, Nucleic Acids Res..

[31]  V. Torre,et al.  Ionic movements through light‐sensitive channels of toad rods. , 1983, The Journal of physiology.

[32]  R. Doolittle,et al.  A simple method for displaying the hydropathic character of a protein. , 1982, Journal of molecular biology.

[33]  W. Rutter,et al.  Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. , 1979, Biochemistry.

[34]  D. Baylor,et al.  The membrane current of single rod outer segments , 1979, Vision Research.

[35]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[36]  R. Kretsinger,et al.  Troponin and parvalbumin calcium binding regions predicted in myosin light chain and T4 lysozyme. , 1975, Science.

[37]  D. Baylor,et al.  Cyclic GMP-activated conductance of retinal photoreceptor cells. , 1989, Annual review of neuroscience.

[38]  T. Weber,et al.  Detection and purification of a recombinant human B lymphotropic virus (HHV-6) in the baculovirus expression system by limiting dilution and DNA dot-blot hybridization. , 1988, Journal of virological methods.

[39]  M. Summers,et al.  A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures. , 1987 .

[40]  R. W. Davis,et al.  Lambda gt 11: gene isolation with antibody probes and other applications. , 1987, Methods in enzymology.

[41]  L. Stryer,et al.  Cyclic GMP cascade of vision. , 1986, Annual review of neuroscience.

[42]  D. Glover.  DNA cloning : a practical approach , 1985 .

[43]  A. Hodgkin,et al.  The ionic selectivity and calcium dependence of the light‐sensitive pathway in toad rods. , 1985, The Journal of physiology.

[44]  C. Yanisch-Perron,et al.  Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. , 1985, Gene.

[45]  J. Messing.  [2] New M13 vectors for cloning , 1983 .