The pathogenic fungus Cryptococcus neoformans produces an extracellular PLB1 (phospholipase B1), shown previously to be a virulence factor. A novel phospholipase (LPL1) with only LPL (lysophospholipase) and LPTA (transacylase) activities has now been characterized in C. gattii, and found to be a 66-kDa glycoprotein (by SDS/PAGE), with a native molecular mass of 670 kDa. The pI was 6.3, and it was active at high temperatures (to 70 °C), as well as at both acidic and neutral pH values. It was stimulated by calcium and palmitoyl carnitine at pH 7.0, but not at pH 5.0, and palmitoyl lysophosphatidylcholine was the preferred substrate. Sequencing indicated that LPL1 is a novel cryptococcal lysophospholipase, and not the gene product of CnLYSO1 or PLB1. A protein with only LPL and LPTA activities was subsequently isolated from two strains of C. neoformans var. grubii. A PLB1 enzyme was isolated from both C. gattii and a highly virulent strain of C. neoformans var. grubii (H99). In both cases, all three enzyme activities (PLB, LPL and LPTA) were present in one 95–120 kDa glycoprotein (by SDS/PAGE) with pI 3.9–4.3. Characterization of PLB1 from C. gattii showed that it differed from that of C. neoformans in its larger native mass (275 kDa), high PLB activity relative to LPL and LPTA, and preference for saturated lipid substrates. Differences in the properties between the secreted phospholipases of the two cryptococcal species could contribute to phenotypic differences that determine their respective environmental niches and different clinical manifestations.
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December 2004
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Research Article|
November 23 2004
Cryptococcal phospholipases: a novel lysophospholipase discovered in the pathogenic fungus Cryptococcus gattii
Lesley C. WRIGHT;
Lesley C. WRIGHT
1
1Centre for Infectious Diseases and Microbiology, University of Sydney at Westmead, Westmead, NSW 2145, Australia, and Department of Infectious Diseases, ICPMR Building, Westmead Hospital, Westmead, NSW 2145, Australia
1To whom correspondence should be addressed (email lesleyw@icpmr.wsahs.nsw.gov.au).
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Jackie PAYNE;
Jackie PAYNE
1Centre for Infectious Diseases and Microbiology, University of Sydney at Westmead, Westmead, NSW 2145, Australia, and Department of Infectious Diseases, ICPMR Building, Westmead Hospital, Westmead, NSW 2145, Australia
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Rosemary T. SANTANGELO;
Rosemary T. SANTANGELO
1Centre for Infectious Diseases and Microbiology, University of Sydney at Westmead, Westmead, NSW 2145, Australia, and Department of Infectious Diseases, ICPMR Building, Westmead Hospital, Westmead, NSW 2145, Australia
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Mukoma F. SIMPANYA;
Mukoma F. SIMPANYA
1Centre for Infectious Diseases and Microbiology, University of Sydney at Westmead, Westmead, NSW 2145, Australia, and Department of Infectious Diseases, ICPMR Building, Westmead Hospital, Westmead, NSW 2145, Australia
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Sharon C. A. CHEN;
Sharon C. A. CHEN
1Centre for Infectious Diseases and Microbiology, University of Sydney at Westmead, Westmead, NSW 2145, Australia, and Department of Infectious Diseases, ICPMR Building, Westmead Hospital, Westmead, NSW 2145, Australia
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Fred WIDMER;
Fred WIDMER
1Centre for Infectious Diseases and Microbiology, University of Sydney at Westmead, Westmead, NSW 2145, Australia, and Department of Infectious Diseases, ICPMR Building, Westmead Hospital, Westmead, NSW 2145, Australia
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Tania C. SORRELL
Tania C. SORRELL
1Centre for Infectious Diseases and Microbiology, University of Sydney at Westmead, Westmead, NSW 2145, Australia, and Department of Infectious Diseases, ICPMR Building, Westmead Hospital, Westmead, NSW 2145, Australia
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Publisher: Portland Press Ltd
Received:
June 24 2004
Revision Received:
August 04 2004
Accepted:
August 23 2004
Accepted Manuscript online:
August 23 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2004
Biochem J (2004) 384 (2): 377–384.
Article history
Received:
June 24 2004
Revision Received:
August 04 2004
Accepted:
August 23 2004
Accepted Manuscript online:
August 23 2004
Citation
Lesley C. WRIGHT, Jackie PAYNE, Rosemary T. SANTANGELO, Mukoma F. SIMPANYA, Sharon C. A. CHEN, Fred WIDMER, Tania C. SORRELL; Cryptococcal phospholipases: a novel lysophospholipase discovered in the pathogenic fungus Cryptococcus gattii. Biochem J 1 December 2004; 384 (2): 377–384. doi: https://doi.org/10.1042/BJ20041079
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