Identification and partial characterization of two proteinases from the cell envelope of Candida albicans blastospores

Rüchel, R.; Böning, Birgitt; Jahn, Edwin C.

doi:10.1016/s0176-6724(85)80068-7

Cited by 16 publications

(10 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2 and 6). Coexistence of more than one extracellular proteinase has also been reported for T. rubrum (3,12,18), Trichophyton mentagrophytes (27,28), and C. albicans (16,17).…”

Section: Discussionmentioning

confidence: 71%

Isolation and properties of extracellular proteinases from Sporothrix schenckii

Tsuboi¹,

T²,

Takamori³

et al. 1987

J Bacteriol

View full text Add to dashboard Cite

Sporothrix schenckii, mainly in the yeast form of the organism, produced extracelluilar proteinases when cultivated in liquid media containing albumin or collagen as a nitrogen source, but did not do so in brain heart infusion medium. Isolation of two extracellular proteinases from albumin-containing medium was performed by chromatography on DEAE-Sepharose CL-6B and Sephacryl S-200. Proteinase I had a molecular weight of 36,500, an optimal pH at 6.0, and a pI at 4.8. Despite its activities in weakly acidic conditions, proteinase I demonstrated chymotrypsinlike characteristics, these being .indicated by strong inhibitory activity by phenylmethylsulfonyl fluoride and chymostatin and good kinetic constants for a synthetic chymotrypsin substrate, Suc-Ala-Ala-Pro-Phe-MCA. Proteinase II had a molecular weight of 39,000, an optimal pH at 3.5, and a pl at 3.8. Proteinase II showed cathepsin D-like characteristics, these being indicated by strong inhibitory activity by pepstatin, an acidic optimal pH, and good kinetic constants for hemoglobin. These two enzymes hydrolyzed natural substrates such as stratum corneum, type I collagen, and elastin although not type IV collagen. Proteinase production and cell growth in collagen-containing medium and the enzymatic digestion of skin constituents by isolated proteinases suggested that these two proteinases cooperatively enable the organism to invade skin and to obtain peptides from insoluble proteins. proteinases, 200-ml samples of BSA-containing media in 500-ml flasks were incubated under the same conditions as described above for 1 week.Assay of proteinase activity. Proteinase activity was assayed with Azocoll (Sigma) as a substrate. A 5-mg sample of Azocoll in 50 mM citric acid-100 mM disodium phosphate buffer was incubated with 50 to 100 ,ul of enzyme solution (total volume, 1.0 ml) for 1 h at 37°C. Incubation was terminated by the addition of 2 ml of cold distilled water. After centrifugation at 1,500 x g for 10 min, released dye in the supernatant was measured at 520 nm with a Hitachi spectrophotometer (model 100-60). One unit of proteinase activity was defined as the amount of enzyme which produced an increase in absorbance of 0.10 under the abovementioned conditions. Determination of protein concentration. Protein concentration was determined by the method of Lowry et al. (9) (Amicon Corp., Lexington, Mass.). After centrifugation at 1,500 x g for 15 min, the supernatant was applied to a DEAE-Sepharose CL-6B column (1.2 by 11 cm) which had been previously equilibrated with the same 4104

show abstract

“…2 and 6). Coexistence of more than one extracellular proteinase has also been reported for T. rubrum (3,12,18), Trichophyton mentagrophytes (27,28), and C. albicans (16,17).…”

Section: Discussionmentioning

confidence: 71%

Isolation and properties of extracellular proteinases from Sporothrix schenckii

Tsuboi¹,

T²,

Takamori³

et al. 1987

J Bacteriol

View full text Add to dashboard Cite

show abstract

“…Others have also found correlations between total proteinase activity and strain virulence (2,7,8). Ruchel et al (18) and Odds (12) studied the ability of different Candida species to produce proteinases and reported variable activity among different Candida species; for example, Cattdida tropicalis produced less proteinase than C. albicans while Candida glabrata does this only occasionally. Other species such as Candida krusei occasionally found in clinical specimens are thought to be non-proteolytic both //; vitro and in vivo (20).…”

mentioning

confidence: 99%

The in vitro proteolytic and saccharolytic activity of Candida species cultured in human saliva

Samaranayake

MacFarlane²,

Samaranayake

et al. 1994

Oral Microbiology and Immunology

View full text Add to dashboard Cite

The proteolytic and saccharolytic activity of 4 Candida species was investigated in batch cultures of pooled, human mixed saliva supplemented with glucose. All the Candida species investigated (Candida albicans, Candida tropicalis, Candida glabrata and Candida krusei) demonstrated a marked growth in saliva with a concomitant reduction in pH from about 7.5 to 3.3, within 72 h. Isotachophoretic analysis of the culture supernatant revealed the presence of a variety of acid anions of which pyruvate and acetate were the most abundant. Proteolysis of salivary components, evaluated by a biochemical assay and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was exhibited by all 4 Candida species, although there was inter-species variation. Despite the similarity in growth rates, C. tropicalis and C. krusei demonstrated greater proteolytic activity than C. albicans and C. glabrata. Neither candidal growth nor proteolysis was observed in glucose-free control saliva samples. In contrast, the degree of saccharolytic and proteolytic activity of a single isolate of C. albicans in glucose-supplemented parotid saliva appeared to be relatively weak compared with mixed saliva. As the oral cavity provides ideal low pH niches periodically supplemented with dietary carbohydrates, the acidic proteinases of Candida species may play a role in the pathogenesis of oral candidiasis.

show abstract

“…In most of the other works, drastic experimental purifying and extracting procedures which could lead to denaturation of the antigenie structure were used [35,36]. With regard to purified molecules of C. albicans cells, with biological activity such as enzymes [37,38], the number of biochemical techniques used for the preparation procedure is usually great [39], leading to a quite low yield of recovery; moreover the purification factor of the molecules is rarely mentioned [40-421. The quantity of 3D9.3 antigen obtained will enable us to develop an immunoenzymatic assay to detect antibodies in humans and in experimental animals, as suggested by Quindos et al [43,44]. We can use this antigen to study the immunological response and it can also be useful for sequencing the epitope.…”

Section: Discussionmentioning

confidence: 99%

Purification of a Candida albicans germ tube specific antigen

Marot-Leblond¹

1995

FEMS Immunology and Medical Microbiology

View full text Add to dashboard Cite

In a previous work, Marot-Leblond et al. identified a Candida albicans germ tube-specific antigen by the use of a monoclonal antibody (mAb 3D9.3). In the present report, we used a two-step procedure to obtain a purified preparation of this antigen from a Zymolyase extract of Candida albicans germ tubes. The extract was first fractionated by gel filtration chromatography. The immunoreactive fractions were pooled, and the 3D9.3 antigen was further purified by hydrophobic interaction chromatography using a Phenyl-superose column. Analysis by SDS-PAGE, immunoblotting and Concanavalin A staining, revealed a single, polydisperse band ranging from 110 to 170 kDa. The antigen was purified 126-fold by protein content and 16.4-fold by carbohydrate content. Recovery of the antigen was 6.8% following the two-step purification.

show abstract

Identification and partial characterization of two proteinases from the cell envelope of Candida albicans blastospores

Cited by 16 publications

References 21 publications

Isolation and properties of extracellular proteinases from Sporothrix schenckii

Isolation and properties of extracellular proteinases from Sporothrix schenckii

The in vitro proteolytic and saccharolytic activity of Candida species cultured in human saliva

Purification of a Candida albicans germ tube specific antigen

Contact Info

Product

Resources

About