Structure of secreted aspartic proteinases from Candida. Implications for the design of antifungal agents.
Pathogens of the genus Candida can cause life threatening infections in immuno-compromised patients. The three-dimensional structures of two closely related secreted aspartic proteinases from C. albicans complexed with a potent (Ki = 0.17 nM) inhibitor, and an analogous enzyme from C. tropicalis reveal variations on the classical aspartic proteinase theme that dramatically alter the specificity of this class of enzymes. The novel fungal proteases present: i) an 8 residue insertion near the first disulfide (Cys45-Cys50, pepsin numbering) that results in a broad flap extending towards the active site; ii) a seven residue deletion replacing helix hN2 (Ser110-Tyr114), which enlarges the S3 pocket; iii) a short polar connection between the two rigid body domains that alters their relative orientation and provides certain specificity; and i.v.) an ordered 12 residue addition at the carboxy terminus. The same inhibitor (A-70450) binds in an extended conformation in the two variants of C. albicans protease, and presents a branched structure at the P3 position. However, the conformation of the terminal methylpiperazine ring is different in the two crystals structures. The implications of these findings for the design of potent antifungal agents are discussed.
Abad-Zapatero
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Laboratory of Protein Crystallography, Abbott Laboratories, Abbott Park, Illinois 60064-3500, USA. abad@abbott.com, abad@mozart.pprd.abbott.com
Goldman
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Muchmore
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Hutchins
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C
Oie
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Stewart
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Cutfield
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Cutfield
J F
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Foundling
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Ray
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D000935:Antifungal Agents / Q000494:pharmacology*; D016282:Aspartic Acid Endopeptidases / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D002175:Candida / Q000187:drug effects / Q000201:enzymology*; D015195:Drug Design; D005656:Fungal Proteins; D006801:Humans; D008958:Models, Molecular; D011480:Protease Inhibitors / Q000737:chemistry; D013379:Substrate Specificity
D000935:Antifungal Agents; D005656:Fungal Proteins; D011480:Protease Inhibitors; D016282:Aspartic Acid Endopeptidases; C086390:SAP1 protein, Candida albicans; C090753:SAP2 protein, Candida