As pigmented structures and fungal surface layer consist mainly of hydrophobic proteins [42], H50 ProteinChip® was chosen in association with

CM10 to compare the profiles obtained from one reference wild-type strain of A. fumigatus (IHEM 18963/Af 293) and four abnormally pigmented strains: three white strains (IHEM 2508, IHEM 9860 and IHEM 13262) 5-Fluoracil and one brown strain (IHEM 15998). Fungal extracts were obtained from three sets of cultures started simultaneously and one set started another day. These cultures were performed on modified Sabouraud medium at 37°C. Since pigments are produced during conidia formation (static culture), we maintained the two oxygenation conditions allowing the analysis of proteins from hyphae and conidia (static culture) and from hyphae (shaken culture). A previous study on these strains [42] has shown that for two of the three white mutants investigated, the ALB1 gene involved early in the melanin synthesis steps

has mutated. For the brown mutant, a point mutation in the ARP2 gene involved in a later step of the Opaganib nmr melanin synthesis has been observed. These three strains presented white or brown powdery colonies. For the strain IHEM 13262, we observed poor conidiation and velvety colonies. As previously observed with the three wild-type strains, the software classified 100% of the metabolic and somatic samples into two clusters in function of oxygenation conditions with the two types of ProteinChips® used (CM10 and H50). Furthermore, the SELDI-TOF-MS analysis of metabolic extracts obtained from static cultures performed on CM10 and on H50 ProteinChips® resulted in the classification of the

five A. fumigatus strains (wild-types and mutants) in five clusters. Figure 3 illustrates the discrimination of the metabolic fractions obtained in static culture from the five strains on CM10 ProteinChip®. Using this ProteinChip® with the five strains under study, eighteen proteins obtained from the metabolic fractions (shaken and static cultures) and thirteen from the somatic extracts (shaken and static cultures) expressed differently (p < 0.05). Some of them were specifically found in the extracts from DCLK1 the wild-type strain in the metabolic and in somatic fractions. On H50 surfaces, only twelve proteins expressed in significantly different ways in the 2 types of extracts. Figure 3 Proteomic comparison between abnormally pigmented strains and a wild-type reference strain of A. fumigatus on CM10 ProteinChips ® . Hierarchical classification of metabolic extracts obtained in static culture for the five strains grown on modified Sabouraud medium at 37°C: white M IHEM 9860 (orange), reference WT strain IHEM 18963 (green), white M IHEM 13262 (red), white M IHEM 2508 (yellow) and brown M IHEM 15998 (blue). The proteins differentially expressed (p < 0.05) were listed on the right of the figure with laser intensities of 2500 nJ (in red) and of 4500 nJ (in blue).