Expression and Characterization of Cryptococcus neoformans Recombinant App1

Asfia Qureshi; Virginia Williams; Maurizio Del Poeta

doi:10.1007/s11046-011-9486-7

Expression and Characterization of Cryptococcus neoformans Recombinant App1

Asfia Qureshi
, Virginia Williams
, Maurizio Del Poeta

Microbiology and Immunology

Medical University of South Carolina

Research output: Contribution to journal › Article › peer-review

6 Scopus citations

Abstract

We characterized Cryptococcus neoformans recombinant antiphagocytic protein 1 (rApp1) by SDS-PAGE, gel filtration chromatography, circular dichroism, and fluorescence spectroscopy. rApp1 produced by C. neoformans var. grubii contains an odd number of cysteines; therefore, it has the ability to form intermolecular disulfide bridges which can lead to the formation of amyloid fibrils in the absence of β-mercaptoethanol or DTT in vitro. Alternate approaches to over-expression of rApp1 in the Lepidopteran High Five™ Insect cell line using pIZ/V5-His and in lentivirus were explored and are described. Finally, localization of App1 in vivo was examined in the presence and absence of the capsule.

Original language	English
Pages (from-to)	395-405
Number of pages	11
Journal	Mycopathologia
Volume	173
Issue number	5-6
DOIs	https://doi.org/10.1007/s11046-011-9486-7
State	Published - Jun 2012

Keywords

Antiphagocytic protein 1 (App1)
Cryptococcus neoformans
Insect cells
Intermolecular disulfide bridges
Lentivirus

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10.1007/s11046-011-9486-7

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title = "Expression and Characterization of Cryptococcus neoformans Recombinant App1",

abstract = "We characterized Cryptococcus neoformans recombinant antiphagocytic protein 1 (rApp1) by SDS-PAGE, gel filtration chromatography, circular dichroism, and fluorescence spectroscopy. rApp1 produced by C. neoformans var. grubii contains an odd number of cysteines; therefore, it has the ability to form intermolecular disulfide bridges which can lead to the formation of amyloid fibrils in the absence of β-mercaptoethanol or DTT in vitro. Alternate approaches to over-expression of rApp1 in the Lepidopteran High Five{\texttrademark} Insect cell line using pIZ/V5-His and in lentivirus were explored and are described. Finally, localization of App1 in vivo was examined in the presence and absence of the capsule.",

keywords = "Antiphagocytic protein 1 (App1), Cryptococcus neoformans, Insect cells, Intermolecular disulfide bridges, Lentivirus",

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AU - Qureshi, Asfia

AU - Williams, Virginia

AU - Del Poeta, Maurizio

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AB - We characterized Cryptococcus neoformans recombinant antiphagocytic protein 1 (rApp1) by SDS-PAGE, gel filtration chromatography, circular dichroism, and fluorescence spectroscopy. rApp1 produced by C. neoformans var. grubii contains an odd number of cysteines; therefore, it has the ability to form intermolecular disulfide bridges which can lead to the formation of amyloid fibrils in the absence of β-mercaptoethanol or DTT in vitro. Alternate approaches to over-expression of rApp1 in the Lepidopteran High Five™ Insect cell line using pIZ/V5-His and in lentivirus were explored and are described. Finally, localization of App1 in vivo was examined in the presence and absence of the capsule.

KW - Antiphagocytic protein 1 (App1)

KW - Cryptococcus neoformans

KW - Insect cells

KW - Intermolecular disulfide bridges

KW - Lentivirus

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M3 - Article

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AN - SCOPUS:84860546862

SN - 0301-486X

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SP - 395

EP - 405

JO - Mycopathologia

JF - Mycopathologia

IS - 5-6

ER -

Expression and Characterization of Cryptococcus neoformans Recombinant App1

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