Rabbit Anti-Human GGH (internal) Polyclonal antibody

The male gamete is not completely mature after ejaculation and requires further events in the female genital tract to acquire fertilizing ability, including the processes of capacitation and acrosome reaction. In order to shed light on protein changes experienced by the sperm cell in preparation for fertilization, a comprehensive quantitative proteomic profiling based on isotopic peptide labeling and liquid chromatography followed by tandem mass spectrometry was performed on spermatozoa from three donors of proven fertility under three sequential conditions: purification with density gradient centrifugation, incubation with capacitation medium, and induction of acrosome reaction by exposure to the calcium ionophore A23187. After applying strict selection criteria for peptide quantification and for statistical analyses, 36 proteins with significant changes in their relative abundance within sperm protein extracts were detected. Moreover, the presence of peptide residues potentially harboring sites for post-translational modification was revealed, suggesting that protein modification may be an important mechanism in sperm maturation. In this regard, increased levels of proteins mainly involved in motility and signaling, both regulated by protein modifiers, were detected in sperm lysates following incubation with capacitation medium. In contrast, less abundant proteins in acrosome-reacted cell lysates did not contain potentially modifiable residues, suggesting the possibility that all those proteins might be relocated or released during the process. Protein-protein interaction analysis revealed a subset of proteins potentially involved in sperm maturation, including the proteins Erlin-2 (ERLIN2), Gamma-glutamyl hydrolase (GGH) and Transmembrane emp24 domain-containing protein 10 (TMED10). These results contribute to the current knowledge of the molecular basis of human fertilization. It should now be possible to further validate the potential role of the detected altered proteins as modulators of male infertility.

Folate derivatives exist in nature in a variety of polyglutamyl forms (Glun); the glutamyl tail is added to the folate molecule by folylpolyglutamate synthetase (FPGS), and removed by gamma-glutamyl hydrolase (GGH) isoforms in several compartments within the cell. Folate polyglutamylation affects the use of the folate cofactors and their transport in organisms, also impacting their bioavailability as vitamins in mammals; however, little is known about its regulation in plants. We explored the possible effect of genetic, developmental, and environmental factors on the Glun extent of the most prevalent folate in plants, 5-CH3-THF. We chose ripening and ethylene treatment in climacteric fruits, to assess developmental and environmental factors. Postharvest ripening increased short Glun tails, and ethylene gassing affected negatively long Glun tails. To evaluate genetic factors, we retrieved and compared the deduced FPGS and GGH sequences from 27 plants with known Glun profile and attempted to correlate their phylogenetic relation, number of isoforms, predicted localization, and primary sequence with the Glun profiles generated and gathered by this study. GGH sequences were quite conserved among plants, while FPGS diverged more. We postulate that the very long Glun tail found only in papaya is the result of very particular changes in one of the FPGS's primary structure.