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Primary structure of a zinc protease from Bacillus mesentericus strain 76
S Stoeva1, T Kleinschmidt, B Mesrob
1Max-Planck-Institut für Biochemie, Abteilung Proteinchemie, Martinsried bei Muenchen, West Germany.
Biochemistry
|January 16, 1990
Summary
The amino acid sequence of Bacillus mesentericus neutral zinc protease (MCP 76) was determined. It is identical to Bacillus subtilis protease, with conserved zinc binding but fewer calcium sites.
Area of Science:
- Biochemistry
- Molecular Biology
- Enzymology
Background:
- Bacillus mesentericus strain 76 produces a neutral zinc protease (MCP 76).
- Understanding protease structure is crucial for enzyme classification and function.
Purpose of the Study:
- To determine the complete amino acid sequence of MCP 76.
- To compare the MCP 76 sequence with other known neutral zinc proteases.
Main Methods:
- Peptide generation using trypsin, chymotrypsin, cyanogen bromide, and o-iodosobenzoic acid.
- Peptide purification via gel filtration and reversed-phase HPLC.
- Amino acid sequence analysis using automatic sequencing.
Main Results:
- The MCP 76 protein comprises 300 amino acid residues.
- The determined sequence is identical to the neutral protease predicted from Bacillus subtilis DNA.
- Conserved residues for zinc and substrate binding were identified.
- A reduced number of calcium binding sites compared to thermolysin was observed.
Conclusions:
- The amino acid sequence provides a basis for classifying MCP 76.
- Structural similarities and differences with other proteases are highlighted, particularly concerning metal ion binding sites.