Expression and sequence of the only detectable aldolase in Chlamydomonas reinhardtii.
Only one aldolase can be resolved from Chlamydomonas reinhardtii by anion-exchange chromatography on DEAE-cellulose. Western blots with specific antisera against plastid and cytosol aldolase of spinach and Northern blots with the respective cDNAs from spinach indicate that only one aldolase, the plastid enzyme, is expressed. Full-length cDNA clones for the plastidic aldolase were isolated from cDNA library constructed from poly(A)+ mRNA of C. reinhardtii with plastid-aldolase-specific probes from spinach. The clones contained a 1.7-kb insert with an open reading frame for 374 amino acid residues covering the mature chloroplast protein of 347 amino acids and a N-terminal transit peptide of 27 amino acids for chloroplast import. The calculated molecular mass of the mature protein is 37.6 kDa and that of the precursor protein is 40.3 kDa. The aldolase of C. reinhardtii shows amino acid similarity of 62 to 67% with the chloroplast aldolase and of 47 to 52% with the cytosolic enzymes of higher plants, respectively. An evolutionary tree with all known class I aldolases shows separate clusters for the chloroplast and for the cytosol aldolases in higher plants and green algae. The aldolase of C. reinhardtii connects at a basal position with the chloroplast aldolases of higher plants.
Schnarrenberger
C
C
Department of Biochemistry, Saga Medical School, Japan.
Pelzer-Reith
B
B
Yatsuki
H
H
Freund
S
S
Jacobshagen
S
S
Hori
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016825:Chlamydomonas reinhardtii / Q000201:enzymology*; D003062:Codon; D018076:DNA, Complementary / Q000235:genetics; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics*; D005796:Genes; D007527:Isoenzymes / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid
D003062:Codon; D018076:DNA, Complementary; D007527:Isoenzymes; D005634:Fructose-Bisphosphate Aldolase