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. 2012 May;26(2):45-9.

Comparison of cell wall proteins in putative Candida albicans & Candida dubliniensis by using modified staining method & SDS-PAGE

Seyed Amir Yazdanparast  1 , Seyedeh Shahrzad Mahdavi Nezarati, Fariba Heshmati, Sepideh Hamzehlou
Affiliations

Comparison of cell wall proteins in putative Candida albicans & Candida dubliniensis by using modified staining method & SDS-PAGE

Seyed Amir Yazdanparast et al. Med J Islam Repub Iran. 2012 May.

Abstract

Background: Candida species are among the most common causes of opportunistic fungal diseases. Among Candida species, Candida albicans is responsible for most infections. Having many strains, C. albicans is very polymorph. C. dubliniensis is very similar to albicans species both morphologically and physiologically. For an infection to occur, cell wall proteins play an important role as they enable yeast to adhere to host cells and begin pathogenesis. Therefore, we decided to extract these proteins and examine them through common molecular methods of protein analysis including SDS-PAGE.

Methods: Initially cell wall proteins of two C. albicans strains (CBS 562 and PTCC6027) and one C. dubliniensis strain (CBS7987) were extracted by using a solution of beta-mercaptoethanol and ammonium carbonate. After dialysis against Tris-HCL buffer, SDS gel electrophoresis was performed on the proteins extract. Bands were then visualized by using three different staining methods among which one method provided improved detection.

Results: By using Coomassie Brilliant Blue staining method, proteins with molecular weight of 42, 66.2 and 200 kDa were detected. By using Silver staining method, proteins with molecular weight of 21.5, 28.5 and 37 kDa were detected. However, using combined Coomassie Brilliant Blue & Sliver staining method visualized more bands resulting in improved detection.

Conclusion: To answer many existing questions about fungal diseases, fungi cell wall proteins are necessary to be examined. To commence such examinations, a simple step may be an SDS-PAGE performance on as many strains as possible. A combined staining method can enhance bands detection.

Keywords: C. dubliniensis; Candida albicans; Protein extraction; SDS-PAGE.

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Figures

Fig. 1
Fig. 1
Candida albicans strain PTCC 6027 (A) Silver stain, (B) Combined staining, (C) Coomassie Brilliant Blue (A & C) The arrowheads showed weak staining bands by a single stain. (B) The upper arrows indicate strong bands by double stain.
Fig. 2
Fig. 2
Candida albicans strain CBS 562 (A) Silver stain, (B) Combined staining, (C) Coomassie Brilliant Blue Asterisks show improved detection of protein bands.
Fig. 3
Fig. 3
Candida Dubliniensis strain CBS 7987 (A/column1, 2 &3) Combined staining, (A/4, 5,6, 7 & 8) Silver staining, (B) Coomassie Brilliant Blue
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