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重组
expressed in E. coli
形式
liquid
比活
≥0.05 U/mg
分子量
19.8 kDa (mature form)
最佳pH
5.0(storage)
7.5(activity)
pI
9.69
运输
dry ice
储存温度
−70°C
一般描述
Alpha-lytic protease (aLP) is an alternative specificity protease for proteomics applications, whose wild-type (WT) version cleaves after T, A, S, and V residues. The M190A (Met190 → Ala190) mutant of aLP has different cleavage specificities, and cleaves after M, F, and L residues. Both the WT and M190A forms of aLP generate peptides of similar average length as trypsin.
In WT aLP, the methionines at positions 190 and 213 cause WT aTP to have its particular specificity toward peptide substrates with small hydrophobic side chains at the P1 position. The M190A mutation gives the resulting mutant M190A aLP the ability to cleave peptide substrates with large hydrophobic side chains at the P1 side chain with greater efficiency compared to WT aLP, in addition to accommodating peptides with small hydrophobic side chains as before. Other structural changes in M190A aLP compared to WT aLP have been discussed.
The activity of M190A aLP in the presence of various solution components is as follows:
In WT aLP, the methionines at positions 190 and 213 cause WT aTP to have its particular specificity toward peptide substrates with small hydrophobic side chains at the P1 position. The M190A mutation gives the resulting mutant M190A aLP the ability to cleave peptide substrates with large hydrophobic side chains at the P1 side chain with greater efficiency compared to WT aLP, in addition to accommodating peptides with small hydrophobic side chains as before. Other structural changes in M190A aLP compared to WT aLP have been discussed.
The activity of M190A aLP in the presence of various solution components is as follows:
- 0.1% sodium deoxycholate: ~1.4-fold enhanced activity
- 1.0% sodium deoxycholate: nearly full activity
- 0.1% SDS: ~40% activity
- 1.0% SDS: ~30% activity
- 1 M guanidine HCl: ~20% activity
- 4 M guanidine HCl: ~1% activity (essentially inactivated)
单位定义
mmols of pNA produced per min per mg protein from 0.5 mM N-succinyl-Ala-Ala-Pro-Phe-PNA at 25 °C in 100 mM Tris-HCl (pH 7.5).
WGK
WGK 1
闪点(°F)
Not applicable
闪点(°C)
Not applicable
法规信息
新产品
Journal of molecular biology, 254(4), 720-736 (1995-12-08)
Gly216 in the active site of the broadly specific MA190 mutant of alpha-lytic protease has been found to be remarkably tolerant of amino acid substitutions. Side-chains as large as Trp can be accommodated within the substrate-binding pocket without abolishing catalysis
Nature, 339(6221), 191-195 (1989-05-18)
The substrate specificity of alpha-lytic protease has been changed dramatically, with a concomitant increase in activity, by replacing an active-site Met with Ala. The substrate specificity of both this mutant and another similar mutant are extraordinarily broad. X-ray crystallographic analysis
Journal of molecular biology, 286(1), 267-278 (1999-02-05)
We have used alpha-lytic protease as a model system for exploring the relationship between the internal dynamics of an enzyme and its substrate specificity. The wild-type enzyme is highly specific for small substrates in its primary specificity pocket, while the
The α-Lytic Protease.
Methods in Enzymology, 19, 599-613 (1970)
Molecular & cellular proteomics : MCP, 13(3), 823-835 (2014-01-16)
Bottom-up proteomics studies traditionally involve proteome digestion with a single protease, trypsin. However, trypsin alone does not generate peptides that encompass the entire proteome. Alternative proteases have been explored, but most have specificity for charged amino acid side chains. Therefore
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