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Chapter 4 : Biochemistry

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Abstract:

This chapter surveys the available information on biochemistry. The capsule is important for virulence and, as a result, the capsular polysaccharide has been studied extensively. Glucuronoxylomannan (GXM), galactoxylomannan (GalXM), and mannoprotein (MP), the three major components of the capsular exopolysaccharides, can each elicit antibody responses, but only the MP component elicits cell-mediated immunity as measured by delayed-type hypersensitivity reaction. The immune response to polysaccharide antigens is discussed. The detection and analysis of capsular polysaccharides in tissue remain dependent upon serological assays. Many assays have been described for the measurement of cryptococcal capsular polysaccharides based on the use of antibody reagents. Electron microscopy of the process of cell wall digestion with snail gut enzymes shows that protoplast-spheroplast formation is a two-stage process. First, the enzymes induce a hole in the equatorial region of the cell wall, through which the protoplast-spheroplast emerges from a cell wall “ ghost”. Second, continued digestion of the cell wall leads to the disappearance of these structures. The biochemistry of melanin and the assembly of melanin on the cell wall remain poorly understood. Melanogenesis is interesting because of its association with virulence and because it is a potential target for antifungal drug design. Mouse passage of environmental isolates produced isolates with higher amphotericin B and fluconazole MICs, suggesting that sterols and antifungal drug resistance could be altered by mammalian infection without a need for exposure to antifungal drugs.

Citation: Casadevall A, Perfect J. 1998. Biochemistry, p 71-114. In . ASM Press, Washington, DC. doi: 10.1128/9781555818241.ch4

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Figure 1

Scanning electron micrograph of encapsulated (top) and nonencapsulated (bottom) strains. On encapsulated strains the capsule appears as a loose fibrillar network (top). For the nonencapsulated strain Cap 67, the surface appears relatively smooth (bottom). Note that bud scars are apparent on some of the nonencapsulated yeast cells. Top micrograph provided by Wendy Cleare (Albert Einstein College of Medicine, Bronx, N.Y.).

Citation: Casadevall A, Perfect J. 1998. Biochemistry, p 71-114. In . ASM Press, Washington, DC. doi: 10.1128/9781555818241.ch4
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Figure 2

Triads identified in GXMs of by proton NMR spectroscopy. Each strain contains a variable amount of each triad (range 0 to 100%). Figure courtesy of Robert Cherniak and reproduced with permission.

Citation: Casadevall A, Perfect J. 1998. Biochemistry, p 71-114. In . ASM Press, Washington, DC. doi: 10.1128/9781555818241.ch4
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Figure 3

Proposed pathway for the oxidation of L-dopa to melanin (reprinted from reference ). Some modifications to this pathway were suggested by Polacheck and Kwon-Chung ( ).

Citation: Casadevall A, Perfect J. 1998. Biochemistry, p 71-114. In . ASM Press, Washington, DC. doi: 10.1128/9781555818241.ch4
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Figure 4

(A) Scanning electron micrograph of melanin ghosts (×15,000). (B) Transmission electron micrograph of a melanin ghost (×45,000). The structure represents melanin left after detergent treatment with guanidinium isothiocyanate and digestion with hot concentrated HCl acid. Reprinted from reference .

Citation: Casadevall A, Perfect J. 1998. Biochemistry, p 71-114. In . ASM Press, Washington, DC. doi: 10.1128/9781555818241.ch4
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Figure 5

Acid phosphatase in cells. Electron-dense deposits inside the cell are due to lead phosphate staining, which indicates the presence of acid phosphatase (×4,500). Note also staining at the edge of the capsule. Figure courtesy of Marta Feldmesser and Phyllis Novikoff (Albert Einstein College of Medicine, Bronx, N.Y.).

Citation: Casadevall A, Perfect J. 1998. Biochemistry, p 71-114. In . ASM Press, Washington, DC. doi: 10.1128/9781555818241.ch4
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References

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Tables

Generic image for table
Table 1

Substances described in culture supernatants

Citation: Casadevall A, Perfect J. 1998. Biochemistry, p 71-114. In . ASM Press, Washington, DC. doi: 10.1128/9781555818241.ch4
Generic image for table
Table 2

Antigenic patterns of serotypes

Based on the factor sera classification of Ikeda et al. ( ).

Citation: Casadevall A, Perfect J. 1998. Biochemistry, p 71-114. In . ASM Press, Washington, DC. doi: 10.1128/9781555818241.ch4
Generic image for table
Table 3

Assays for the serological detection of cryptococcal polysaccharide

Citation: Casadevall A, Perfect J. 1998. Biochemistry, p 71-114. In . ASM Press, Washington, DC. doi: 10.1128/9781555818241.ch4
Generic image for table
Table 4

Biological phenomena associated with and attributed to melanin production in

Citation: Casadevall A, Perfect J. 1998. Biochemistry, p 71-114. In . ASM Press, Washington, DC. doi: 10.1128/9781555818241.ch4
Generic image for table
Table 5

Some enzymes and enzyme activities and their cellular location in

Location refers to site of isolation of enzyme or enzyme activity according to the reference cited.

Citation: Casadevall A, Perfect J. 1998. Biochemistry, p 71-114. In . ASM Press, Washington, DC. doi: 10.1128/9781555818241.ch4
Generic image for table
Table 6

Sterol composition of 13 isolates in the presence or absence of fluconazole

Adopted from reference . Values are means ± standard deviations. values are obtained by -test and adjusted for the Bonferroni correction.

Citation: Casadevall A, Perfect J. 1998. Biochemistry, p 71-114. In . ASM Press, Washington, DC. doi: 10.1128/9781555818241.ch4

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