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Comparison of cell wall proteins in putative Candida albicans & Candida dubliniensis by using modified staining
Seyed Amir Yazdanparast1, Seyedeh Shahrzad Mahdavi Nezarati, Fariba Heshmati
1PhD, Associate Professor, Department of Medical Mycology, School of Allied Health Sciences, Tehran University of Medical Sciences, Shahid Hemmat Expressway, Shahid Chamran Crossway, Tehran, Iran.
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.
Insights
Investigating Candida species cell wall proteins using SDS-PAGE revealed distinct molecular weights. A combined staining method enhanced detection, crucial for understanding fungal pathogenesis.
Area of Science:
- Mycology
- Molecular Biology
- Biochemistry
Background:
- Candida species are common opportunistic fungal pathogens, with Candida albicans causing most infections.
- Candida albicans exhibits significant strain polymorphism, and Candida dubliniensis shares similarities.
- Cell wall proteins are crucial for Candida adhesion and pathogenesis.
Purpose of the Study:
- To extract and analyze cell wall proteins from Candida species.
- To compare protein profiles of different Candida strains using molecular methods.
- To optimize protein detection methods for Candida cell wall analysis.
Main Methods:
- Extraction of cell wall proteins from Candida albicans and Candida dubliniensis strains.
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for protein separation.
- Visualization of protein bands using Coomassie Brilliant Blue, Silver staining, and a combined method.
Main Results:
- Coomassie Brilliant Blue staining detected proteins at 42, 66.2, and 200 kDa.
- Silver staining identified proteins at 21.5, 28.5, and 37 kDa.
- A combined Coomassie Brilliant Blue and Silver staining method improved band visualization and detection.
Conclusions:
- Examination of fungal cell wall proteins is essential for understanding fungal diseases.
- SDS-PAGE is a valuable technique for analyzing Candida cell wall proteins across multiple strains.
- Combined staining protocols enhance the detection of fungal cell wall proteins.
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