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PDBsum entry 3fzd
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References listed in PDB file
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Key reference
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Title
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Mutation of asn28 disrupts the dimerization and enzymatic activity of sars 3cl(pro) .
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Authors
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J.Barrila,
S.B.Gabelli,
U.Bacha,
L.M.Amzel,
E.Freire.
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Ref.
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Biochemistry, 2010,
49,
4308-4317.
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PubMed id
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Abstract
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Coronaviruses are responsible for a significant proportion of annual respiratory
and enteric infections in humans and other mammals. The most prominent of these
viruses is the severe acute respiratory syndrome coronavirus (SARS-CoV) which
causes acute respiratory and gastrointestinal infection in humans. The
coronavirus main protease, 3CL(pro), is a key target for broad-spectrum
antiviral development because of its critical role in viral maturation and high
degree of structural conservation among coronaviruses. Dimerization is an
indispensable requirement for the function of SARS 3CL(pro) and is regulated
through mechanisms involving both direct and long-range interactions in the
enzyme. While many of the binding interactions at the dimerization interface
have been extensively studied, those that are important for long-range control
are not well-understood. Characterization of these dimerization mechanisms is
important for the structure-based design of new treatments targeting
coronavirus-based infections. Here we report that Asn28, a residue 11 A from the
closest residue in the opposing monomer, is essential for the enzymatic activity
and dimerization of SARS 3CL(pro). Mutation of this residue to alanine almost
completely inactivates the enzyme and results in a 19.2-fold decrease in the
dimerization K(d). The crystallographic structure of the N28A mutant determined
at 2.35 A resolution reveals the critical role of Asn28 in maintaining the
structural integrity of the active site and in orienting key residues involved
in binding at the dimer interface and substrate catalysis. These findings
provide deeper insight into complex mechanisms regulating the activity and
dimerization of SARS 3CL(pro).
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