Comparative Analysis and Ancestral Sequence Reconstruction of Bacterial Sortase Family Proteins Generates Functional Ancestral Mutants with Different Sequence Specificities
نویسندگان
چکیده
Gram-positive bacteria are some of the earliest known life forms, diverging from gram-negative 2 billion years ago. These organisms utilize sortase enzymes to attach proteins their peptidoglycan cell wall, a structural feature that distinguishes two types bacteria. The transpeptidase activity sortases make them an important tool in protein engineering applications, e.g., sortase-mediated ligations or sortagging. However, due relatively low catalytic efficiency, there ongoing efforts create better variants for these uses. Here, we use bioinformatics tools, principal component analysis and ancestral sequence reconstruction, combination with biochemistry, analyze natural variation enzymes. Principal on superfamily previously described classes identifies regions high structurally-conserved loops within each family, including those near active site. Using determined sequences Staphylococcus Streptococcus Class A proteins. Enzyme assays revealed enzyme is shares similar other sortases. Taken together, highlight how can be utilized investigate this arguing techniques may used discover design increased efficiency and/or selectivity ligation experiments.
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ژورنال
عنوان ژورنال: Bacteria
سال: 2022
ISSN: ['2674-1334']
DOI: https://doi.org/10.3390/bacteria1020011