Close evolutionary relatedness among functionally distantly related members of the (alpha/beta)8-barrel glycosyl hydrolases suggested by the similarity of their fifth conserved sequence region.
A short conserved sequence equivalent to the fifth conserved sequence region of alpha-amylases (173_LPDLD, Aspergillus oryzae alpha-amylase) comprising the calcium-ligand aspartate, Asp-175, was identified in the amino acid sequences of several members of the family of (alpha/beta)8-barrel glycosyl hydrolases. Despite the fact that the aspartate is not invariantly conserved, the stretch can be easily recognised in all sequences to be positioned 26-28 amino acid residues in front of the well-known catalytic aspartate (Asp-206, A. oryzae alpha-amylase) located in the beta 4-strand of the barrel. The identification of this region revealed remarkable similarities between some alpha-amylases (those from Bacillus megaterium, Bacillus subtilis and Dictyoglomus thermophilum) on the one hand and several different enzyme specificities (such as oligo-1,6-glucosidase, amylomaltase and neopullulanase, respectively) on the other hand. The most interesting example was offered by B. subtilis alpha-amylase and potato amylomaltase with the regions LYDWN and LYDWK, respectively. These observations support the idea that all members of the family of glycosyl hydrolases adopting the structure of the alpha-amylase-type (alpha/beta)8-barrel are mutually closely related and the strict evolutionary borders separating the individual enzyme specificities can be hardly defined.
Institute of Ecobiology, Slovak Academy of Sciences, Bratislava, Slovakia.
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Janecek
D000595:Amino Acid Sequence; D001410:Bacillus megaterium / Q000201:enzymology; D001412:Bacillus subtilis / Q000201:enzymology; D005075:Biological Evolution; D017124:Conserved Sequence; D006004:Glycogen Debranching Enzyme System / Q000737:chemistry; D006026:Glycoside Hydrolases / Q000737:chemistry*; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D017385:Sequence Homology; D011198:Solanum tuberosum / Q000201:enzymology; D000516:alpha-Amylases / Q000737:chemistry
D006004:Glycogen Debranching Enzyme System; C020569:4 alpha-glucanotransferase; D006026:Glycoside Hydrolases; D000516:alpha-Amylases