Thermal stability of homologous neutral metalloendopeptidases in thermophilic and mesophilic bacteria: structural considerations.
Thermolysin and neutral protease A are neutral metalloendopeptidases having similar specificity, molecular weight, metal content, and amino acid composition. Thermolysin, derived from the thermophilic organism Bacillus thermoproteolyticus, is heat inactivated at about 84 degrees whereas neutral protease A, derived from the mesophilic organism Bacillus subtilis, is inactivated at about 59 degrees. Structural analyses reveal that the two enzymes are homologous. Of the 326 residues of neutral protease A, 171 have been placed in sequence and 49% of these have been found in identical loci in thermolysin. These include many of the residues corresponding to the active site of thermolysin. The sensitivity of both enzymes to thermal inactivation is dependent upon the presence of calcium and neutral protease appears to bind less calcium than thermolysin. Structural data indicate that many of the ligands associated with calcium sites 1 and 2 (double site of thermolysin) are present in neutral protease and that calcium site 4 cannot exist in neutral protease. The structural homology and functional analogy of these two proteins support the concept that they have similar conformations. The known structure of thermolysin is used as a model to discuss structural differences which might be related to thermal stability.
D000595:Amino Acid Sequence; D000812:Anhydrides; D001407:Bacillus / Q000201:enzymology*; D001412:Bacillus subtilis / Q000201:enzymology; D002118:Calcium / Q000032:analysis; D005561:Formates / Q000494:pharmacology; D006358:Hot Temperature; D008628:Mercury / Q000494:pharmacology; D008667:Metalloproteins / Q000032:analysis; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000032:analysis* / Q000378:metabolism; D011487:Protein Conformation; D011489:Protein Denaturation; D012834:Silver / Q000494:pharmacology; D013820:Thermolysin / Q000032:analysis* / Q000378:metabolism; D015032:Zinc / Q000032:analysis
D000812:Anhydrides; D005561:Formates; D008667:Metalloproteins; D012834:Silver; D010447:Peptide Hydrolases; D013820:Thermolysin; D008628:Mercury; D015032:Zinc; D002118:Calcium