Background and Objective Gingival crevicular fluid (GCF) has been of major interest for many decades as valuable body fluid that may serve as a source of biomarkers for both periodontal and systemic diseases. Because of its very small sample size, sub-μl level, identification of its protein composition by classical biochemical methods has been limited. The advent of highly sensitive mass spectrometric technology has permitted large-scale identification of protein components of many biological samples. This technology has been employed to identify protein composition of GCF from inflamed and periodontal sites. In this report we present a proteome dataset of GCF from healthy periodontium sites. Methods A combination of periopaper collection method with application of multidimensional protein separation and mass spectrometric (MS) technology led to a large-scale documentation of the proteome of GCF from healthy periodontium sites. Results The approaches utilized have culminated in identification of 199 proteins in GCF of periodontally healthy sites. The current GCF proteome from healthy sites was compared and contrasted with those proteomes of GCF from inflamed and periodontal sites as well as serum. The cross-correlation of the GCF and plasma proteomes permitted dissociation of the 199 identified GCF proteins into, 105 proteins (57%) that can be identified in plasma and 94 proteins (43%) which are distinct and unique to GCF microenvironment. Such analysis also revealed distinctions in protein functional categories between serum proteins and those specific to GCF microenvironment. Conclusion Firstly, the data presented herein provide the proteome of GCF from periodontally healthy sites through establishment of innovative analytical approaches for effective analysis of GCF from periopapers both at the level of complete elusion and removal of abundant albumin which restricts identification of low abundant proteins. Secondly, it adds significantly to the knowledge of GCF composition and highlights new groups of proteins specific to GCF microenvironment.

Naturally occurring salivary antifungal proteins have been of major interest, due to their potential to provide the basis for peptide antimycotics effective in combating fungal infections in the oral cavity or elsewhere. We tested the fungistatic activity of a number of cationic antifungal proteins, with major emphasis on histatin 5, a basic protein secreted by the human parotid and submandibular glands. Histatin 5 inhibited the growth of Candida albicans and that of other medically important Candida species, such as C. kefyr, C. krusei, and C. parapsilosis, with IC 50 values in the range of 10-20 µg/ml. Two Cryptococcus neoformans strains were also sensitive (IC 50 5.2 and 5.6 µg/ml). On the other hand, three C. glabrata strains (ATCC 90030, 2001 and 64677) were entirely insensitive to histatin 5 (IC 50 >225 µg/ml). Four genetically very similar species to C. glabrata, Candida castelli (CBS 4332), Saccharomyces cerevisiae (S288C, BY4741 and CBS 1171), Kluyveromyces delphensis (CBS 2170) and Kluyveromyces bacillisporus (CBS 7720) were all sensitive to histatin 5 (IC 50 2.6-64.6 µg/ml). C. glabrata was also insensitive to other members of the histatin family; histatin 1, 3 and P-113 (IC 50 values in all cases >225 µg/ml). In addition, two entirely different cationic antifungal proteins originating from frog skin, PGLa and magainin 2, also showed a strong reduced activity toward this fungus. Besides the well-described inherent resistance of C. glabrata to azole-derived antifungal agents, our studies indicate that this species is also able to withstand the otherwise detrimental activities of cationic antifungal proteins.

Histatin 5, a human salivary protein with broad-spectrum antifungal activity, is remarkably ineffective against Candida glabrata. Fluconazole resistance in this fungus is due in most cases to upregulation of CgCDR efflux pumps. We investigated whether the distinct resistance of C. glabrata to histatin 5 is related to similar mechanisms.