Sequence of the Kluyveromyces lactis beta-galactosidase: comparison with prokaryotic enzymes and secondary structure analysis.
The LAC4 gene encoding the beta-galactosidase (beta Gal) of the yeast, Kluyveromyces lactis, was cloned on a 7.2-kb fragment by complementation of a lacZ-deficient Escherichia coli strain. The nucleotide sequence of the structural gene, with 42 bp and 583 bp of the 5'- and 3'-flanking sequences, respectively, was determined. The deduced amino acid (aa) sequence of the K. lactis beta Gal predicts a 1025-aa polypeptide with a calculated M(r) of 117618 and reveals extended sequence homologies with all the published prokaryotic beta Gal sequences. This suggests that the eukaryotic beta Gal is closely related, evolutionarily and structurally, to the prokaryotic beta Gal's. In addition, sequence similarities were observed between the highly conserved N-terminal two-thirds of the beta Gal and the entire length of the beta-glucuronidase (beta Glu) polypeptides, which suggests that beta Glu is clearly related, structurally and evolutionarily, to the N-terminal two-thirds of the beta Gal. The structural analysis of the beta Gal alignment, performed by mean secondary structure prediction, revealed that most of the invariant residues are located in turn or loop structures. The location of the invariant residues is discussed with respect to their accessibility and their possible involvement in the catalytic process.
Poch
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O
Institut de Biologie Moléculaire et Cellulaire du CNRS, Strasbourg, France.
L'Hôte
H
H
Dallery
V
V
Debeaux
F
F
Fleer
R
R
Sodoyer
R
R
D000595:Amino Acid Sequence; D001483:Base Sequence; D005075:Biological Evolution; D003001:Cloning, Molecular; D005057:Eukaryotic Cells; D005800:Genes, Fungal / Q000235:genetics; D014644:Genetic Variation; D005966:Glucuronidase / Q000235:genetics*; D007716:Kluyveromyces / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D011387:Prokaryotic Cells; D011487:Protein Conformation; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013329:Structure-Activity Relationship; D001616:beta-Galactosidase / Q000235:genetics*
D001616:beta-Galactosidase; D005966:Glucuronidase