Global Proteomic Profiling of the Secretome of Candida albicans ecm33 Cell Wall Mutant Reveals the Involvement of Ecm33 in Sap2 Secretion.
Candida albicans secretes numerous proteins related to cell wall remodeling, adhesion, nutrient acquisition and host interactions. Also, extracellular vesicles containing cytoplasmic proteins are secreted into the medium. The C. albicans ecm33/ecm33 mutant (RML2U) presents an altered cell wall and is avirulent. The proteomic analysis of proteins secreted by RML2U cells identified a total of 170 proteins: 114 and 154 of which correspond to the vesicle-free secretome and extracellular vesicles, respectively. Notably, 98 proteins were common to both samples, and the groups most represented were metabolic and cell wall-related proteins. The results of this study showed that RML2U had an altered pattern of proteins secreted by the classical secretion pathway as well as the formation of extracellular vesicles, including their size, quantity, and protein composition. Specifically, the secretion of aspartic protease 2 (Sap2) was compromised but not its intracellular expression, with bovine serum albumin (BSA) degradation by RML2U being altered when BSA was used as the sole nitrogen source. Furthermore, as recent research links the expression of Sap2 to the TOR (Target Of Rapamycin) signaling pathway, the sensitivity of RML2U to rapamycin (the inhibitor of TOR kinase) was tested and found to be enhanced, connecting Ecm33 with this pathway.
Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense de Madrid , Plaza de Ramón y Cajal s/n, 28040 Madrid, Spain.|Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS) , Ctra. de Colmenar Viejo, 28034 Madrid, Spain.
Ana
A
Gil-Bona
Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense de Madrid , Plaza de Ramón y Cajal s/n, 28040 Madrid, Spain.|Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS) , Ctra. de Colmenar Viejo, 28034 Madrid, Spain.
Lucía
L
Monteoliva
Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense de Madrid , Plaza de Ramón y Cajal s/n, 28040 Madrid, Spain.|Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS) , Ctra. de Colmenar Viejo, 28034 Madrid, Spain.
Concha
C
Gil
D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D016282:Aspartic Acid Endopeptidases / Q000235:genetics* / Q000378:metabolism; D002176:Candida albicans / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D002473:Cell Wall / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D000067128:Extracellular Vesicles / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D059748:Proteolysis; D040901:Proteomics / Q000379:methods; D012710:Serum Albumin, Bovine / Q000737:chemistry; D020123:Sirolimus / Q000494:pharmacology; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
D000935:Antifungal Agents; D005656:Fungal Proteins; D008565:Membrane Proteins; D012710:Serum Albumin, Bovine; D058570:TOR Serine-Threonine Kinases; D016282:Aspartic Acid Endopeptidases; C090753:SAP2 protein, Candida; D020123:Sirolimus
Candida albicans; ECM33; LC−MS/MS analysis; extracellular vesicles; rapamycin; secreted aspartyl protease (Sap2); secreted proteins