Molecular cloning and sequence analysis of a cDNA encoding a porcine kidney renin-binding protein.
A complementary DNA encoding a renin-binding protein (RnBP) has been isolated from a porcine kidney cDNA library by immunological screening of in vitro translation products from the cDNAs. Analysis of the nucleotide sequence of the clone revealed a 1,342-nucleotide sequence with a 5'-terminal untranslated region of 52 nucleotides, an open reading frame of 1,206 nucleotides that encodes 402 amino acids, and a 3'-terminal untranslated region of 84 nucleotides that contains the polyadenylation signal sequence, AATAAA. The predicted amino acid sequence contains no hydrophobic amino-terminal sequence and does not show significant homology to those of other identified proteins. The in vitro translated RnBP was found to have the same molecular weight, 42,000, as that of the purified RnBP from porcine kidney on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and it formed a complex with renin purified from porcine kidney, which indicates that the cDNA encodes a functional RnBP without a propeptide sequence. The RnBP cDNA probe hybridized to a 1.5-kilobase mRNA in kidney, liver, adrenal, and pituitary glands, the amount being much greater in kidney than in the other tissues. Southern blot analysis showed the presence of a unique gene for RnBP in the porcine genome.
Inoue
H
H
Department of Biochemistry, National Cardiovascular Center Research Institute, Osaka, Japan.
Fukui
K
K
Takahashi
S
S
Miyake
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002238:Carbohydrate Epimerases; D002352:Carrier Proteins / Q000235:genetics* / Q000302:isolation & purification; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D015723:Gene Library; D007668:Kidney / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D014176:Protein Biosynthesis; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000302:isolation & purification; D012083:Renin / Q000378:metabolism*; D015183:Restriction Mapping; D013552:Swine
D002352:Carrier Proteins; D011994:Recombinant Proteins; D004247:DNA; D012083:Renin; D002238:Carbohydrate Epimerases; C024519:N-acyl-D-glucosamine 2-epimerase