Characterization of Recombinant Terrelysin,a Hemolysin of <Emphasis Type="Italic">Aspergillus terreus</Emphasis> |
| |
| Authors: | Ajay P Nayak Françoise M Blachere Justin M Hettick Slawomir Lukomski Detlef Schmechel Donald H Beezhold |
| |
| Institution: | (1) Allergy and Clinical Immunology Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, 1095 Willowdale Rd, Morgantown, WV, 26505, USA;(2) Department of Microbiology, Immunology and Cell Biology, West Virginia University, Morgantown, WV 26505, USA; |
| |
| Abstract: | Fungal hemolysins are potential virulence factors. Some fungal hemolysins belong to the aegerolysin protein family that includes
cytolysins capable of lysing erythrocytes and other cells. Here, we describe a hemolysin from Aspergillus terreus called terrelysin. We used the genome sequence database to identify the terrelysin sequence based on homology with other
known aegerolysins. Aspergillus terreus mRNA was isolated, transcribed to cDNA and the open reading frame for terrelysin amplified by PCR using specific primers.
Using the pASK-IBA6 cloning vector, we produced recombinant terrelysin (rTerrelysin) as a fusion product in Escherichia coli. The recombinant protein was purified and using MALDI-TOF MS determined to have a mass of 16,428 Da. Circular dichroism analysis
suggests the secondary structure of the protein to be predominantly β-sheet. Results from thermal denaturation of rTerrelysin
show that the protein maintained the β-sheet confirmation up to 65°C. Polyclonal antibody to rTerrelysin recognized a protein
of approximately 16.5 kDa in mycelial extracts from A. terreus. |
| |
| Keywords: | |
| 本文献已被 SpringerLink 等数据库收录！ |
|
