An essential tyrosine residue of Aspergillus polygalacturonase.
Based on strict conservation of a tyrosine residue in 24 polygalacturonases, tyrosine modification was assessed in two different forms of the Aspergillus enzyme. The second subform was unknown in structure but submitted to sequence analysis and was found also to have the conserved tyrosine residue. Results of chemical modifications are consistent in showing inactivation of the proteins with all tyrosine-reactive agents tested, acetic anhydride, N-acetyl imidazole, and tetranitromethane. Furthermore, after acetylation, regeneration of enzyme activity was possible with hydroxylamine. Spectrophotometric pH titration showed that one accessible tyrosine residue is ionized at pH 9.3-9.5, whereas the remaining, masked residues are all ionized at pH 10.5. It is concluded that one tyrosine residue is catalytically important, in agreement with the inactivation and reactivation data, that this residue is accessible, and that it is likely to correspond to the strictly conserved residue observed in all forms.
Stratilová
E
E
Institute of Chemistry, Slovak Academy of Sciences, Bratislava, Slovakia.
Dzúrová
M
M
Markovic
O
O
Jörnvall
H
H
D000089:Acetic Anhydrides / Q000494:pharmacology; D000107:Acetylation; D000595:Amino Acid Sequence; D001230:Aspergillus / Q000201:enzymology*; D004789:Enzyme Activation; D004793:Enzyme Reactivators / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D019811:Hydroxylamine; D006898:Hydroxylamines / Q000494:pharmacology; D007093:Imidazoles / Q000494:pharmacology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D011096:Polygalacturonase / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D017421:Sequence Analysis; D013774:Tetranitromethane / Q000494:pharmacology; D014443:Tyrosine / Q000032:analysis / Q000502:physiology*
D000089:Acetic Anhydrides; D004793:Enzyme Reactivators; D006898:Hydroxylamines; D007093:Imidazoles; D010446:Peptide Fragments; C031800:acetic anhydride; D019811:Hydroxylamine; D014443:Tyrosine; D011096:Polygalacturonase; D013774:Tetranitromethane; C012160:N-acetylimidazole