title: Identification of fibrinogen-binding proteins of Aspergillus fumigatus using proteomic approach.
abstract: Aspergillus fumigatus, the main etiological agent for various forms of human aspergillosis, gets access to the respiratory system of human host by inhalation of airborne conidia. These conidia possibly adhere to extracellular matrix (ECM) proteins. Among the ECM proteins involved in adherence, fibrinogen is thought to be crucial. Here, we studied whether A. fumigatus three-week culture filtrate (3wcf) proteins promote binding of A. fumigatus to ECM proteins and promote fungal growth. We observed that incubation of ECM with 3wcf proteins led to dose- and time-dependent increase in adherence of conidia to the ECM. In order to identify the catalogue of fibrinogen-binding A. fumigatus proteins, we carried out fibrinogen affinity blotting using two-dimensional gel electrophoresed 3wcf proteins. A total of 15 fibrinogen-binding protein spots corresponding to 7 unique proteins were identified in 3wcf using matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF-TOF). Among these, 4 proteins, namely, beta-glucosidase, alpha-mannosidase, pectate lyase A and oryzin precursor were predicted to have cell wall or extracellular localization, whereas amidase family protein and two hypothetical proteins did not display the signal sequence. This study reports seven novel fibrinogen-binding proteins of A. fumigatus, some of which could be further explored for targeting the adhesion phenomenon as antifungal strategy.
authors.affiliation: Institute of Genomics and Integrative Biology, Council for Scientific and Industrial Research, Delhi, India.
authors.forename: Santosh Kumar
authors.initials: SK
authors.lastname: Upadhyay
authors.forename: Poonam
authors.initials: P
authors.lastname: Gautam
authors.forename: Hrishikesh
authors.initials: H
authors.lastname: Pandit
authors.forename: Yogendra
authors.initials: Y
authors.lastname: Singh
authors.forename: Seemi Farhat
authors.initials: SF
authors.lastname: Basir
authors.forename: Taruna
authors.initials: T
authors.lastname: Madan
mesh_terms: D000595:Amino Acid Sequence; D001228:Aspergillosis / Q000378:metabolism* / Q000382:microbiology; D001232:Aspergillus fumigatus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D002352:Carrier Proteins; D016326:Extracellular Matrix Proteins; D005656:Fungal Proteins; D005340:Fibrinogen