Abstract
Cells produce proteases as inactive zymogens. Here, we demonstrate that this tactic can extend beyond proteases. By linking the N and C termini of ribonuclease A, we obstruct the active site with the amino acid sequence recognized by plasmepsin II, a highly specific protease from Plasmodium falciparum. We generate new N and C termini by circular permutation. In the presence of plasmepsin II, a ribonuclease zymogen gains approximately 10(3)-fold in catalytic activity and maintains high conformational stability. We conclude that zymogen creation provides a new and versatile strategy for the control of enzymatic activity, as well as the potential development of chemotherapeutic agents.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Animals
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Aspartic Acid Endopeptidases / metabolism
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Catalytic Domain / genetics
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Cattle
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Enzyme Activation
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Enzyme Precursors / chemistry*
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Enzyme Precursors / genetics
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Enzyme Precursors / metabolism
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Enzyme Stability
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Models, Molecular
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Molecular Sequence Data
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Plasmids / genetics
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Protein Conformation
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Protozoan Proteins
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Ribonuclease, Pancreatic / chemistry*
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Ribonuclease, Pancreatic / genetics
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Ribonuclease, Pancreatic / metabolism
Substances
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Enzyme Precursors
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Protozoan Proteins
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Recombinant Proteins
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Ribonuclease, Pancreatic
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Aspartic Acid Endopeptidases
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plasmepsin II