Spinach carbonic anhydrase primary structure deduced from the sequence of a cDNA clone.
A cDNA clone 1,156 base pairs in length was selected by screening a lambda gt11 library with antibodies directed against spinach chloroplast carbonic anhydrase (carbonate dehydratase, EC 4.2.1.1). Sequence analysis revealed an open reading frame of 957 base pairs encoding a polypeptide containing 319 amino acids with a molecular weight of 34,569. This polypeptide is of sufficient size to represent the precursor of spinach chloroplast carbonic anhydrase. The polypeptide contains a sequence of 19 amino acids identical to the sequence of a cyanogen bromide fragment from spinach carbonic anhydrase. In addition, Escherichia coli was transformed with a plasmid that expresses spinach carbonic anhydrase. Lysates prepared from transformed E. coli contain acetazolamide-inhibitable carbonic anhydrase activity. The amino acid sequence of spinach carbonic anhydrase is distinct from those reported for the mammalian isozymes.
Fawcett
T W
TW
Department of Biochemistry, Louisiana State University, Baton Rouge 70803.
Browse
J A
JA
Volokita
M
M
Bartlett
S G
SG
D000595:Amino Acid Sequence; D001483:Base Sequence; D002256:Carbonic Anhydrases / Q000235:genetics*; D002736:Chloroplasts / Q000201:enzymology; D004247:DNA / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008969:Molecular Sequence Data; D010944:Plants / Q000201:enzymology* / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D014169:Transformation, Bacterial
D004247:DNA; D002256:Carbonic Anhydrases