Proteomic analysis of seminal plasma from infertile patients with oligoasthenoteratozoospermia due to oxidative stress and comparison with fertile volunteers

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Proteomic analysis of seminal plasma from infertile patients with oligoasthenoteratozoospermia due to oxidative stress and comparison with fertile volunteers Ralf Herwig, M.D., Christian Knoll, M.Sc., Melanie Planyavsky, Ali Pourbiabany, Joachim Greilberger, Ph.D., Keiryn L. Bennett, Ph.D. Fertility and Sterility Volume 100, Issue 2, Pages 355-366.e2 (August 2013) DOI: 10.1016/j.fertnstert.2013.03.048 Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 1 An overview of the preparation steps from seminal ejaculate collection and clinical assessment to identification of the proteins by liquid chromatography tandem mass spectrometry (LCMS) and protein database searching. Fertility and Sterility 2013 100, 355-366.e2DOI: (10.1016/j.fertnstert.2013.03.048) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Venn diagram of the number of proteins identified by liquid chromatography tandem mass spectrometry (LCMS) of seminal plasma from (A) all fertile versus all idiopathic oligoasthenoteratozoospermia (iOAT); (B) 11 pooled fertile versus 11 pooled iOAT; (C) single fertile versus single iOAT; and (D) pooled iOAT versus single patient iOAT. Fertility and Sterility 2013 100, 355-366.e2DOI: (10.1016/j.fertnstert.2013.03.048) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Summary of the 27 common proteins identified by liquid chromatography tandem mass spectrometry (LCMS) from all idiopathic oligoasthenoteratozoospermia (iOAT) patients investigated in this study (upper panel) and the 24 proteins identified by LCMS that were ≥1.5-fold more highly expressed in median spectral counts (that is, the number of peptide to spectrum matches) in iOAT patients compared with in fertile control subjects (lower panel). The number of uniquely identified peptides, spectral counts (that is, the number of peptide to spectrum matches), and protein sequence coverage are given for each technical replicate of the 11 pooled samples and the single patient analysis. Proteins are sorted on the basis of the global spectral counts, which is an approximate indicator of the abundance of protein in the samples. Fertility and Sterility 2013 100, 355-366.e2DOI: (10.1016/j.fertnstert.2013.03.048) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 4 GO functional annotation chart of the 46 proteins identified in this study as implicated in idiopathic oligoasthenoteratozoospermia (iOAT) (GOTERM_BP_ALL). Analysis was performed on the 27 proteins identified in common between all iOAT patients investigated (red bars) and the 24 proteins identified as ≥1.5-fold more highly expressed in median spectral counts (that is, the number of peptide to spectrum matches) in iOAT patients compared with in fertile control subjects (blue bars). The DAVID bioinformatic resources tool was used for the analysis with Count and EASE thresholds of 2 (default) and 0.5, respectively. Fertility and Sterility 2013 100, 355-366.e2DOI: (10.1016/j.fertnstert.2013.03.048) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Box and whiskers plot for (A) total protein concentration and (B) carbonyl protein (CP) levels. Fertility and Sterility 2013 100, 355-366.e2DOI: (10.1016/j.fertnstert.2013.03.048) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 2 BIOCARTA and KEGG pathway analysis of the proteins identified in the single fertile and iOAT samples (P<.05). The DAVID bioinformatic resources tool was used for the analysis with Count and EASE thresholds of 2 (default) and 0.1 (default), respectively. Pathways enriched are given for fertile only (green bars), common to fertile and iOAT (red bars), and iOAT only (blue bars). Fertility and Sterility 2013 100, 355-366.e2DOI: (10.1016/j.fertnstert.2013.03.048) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions