This study aimed to develop protocols for the extraction of sperm proteins from Moxotó goats (Capra hircus) and to compare the resulting proteomic maps. The sperm proteins were isolated using an extraction buffer containing 7 M urea and 2 M thiourea, 20 mM DTT, and one of the following detergents: 1% or 4% CHAPS; 1% or 4% SDS; 1% or 4% Triton X-100; or a combination of CHAPS and SDS. The 1-DE and 2-DE profiles of the isolated proteins revealed that the various isolation methods were efficient. Qualitative and quantitative differences in the 1-DE and 2-DE profiles were observed. 2-DE maps indicated that the amount and diversity of proteins visualized depended on the detergent that was used. Furthermore, this work revealed that the combination of detergents increased the resolution of some spots and retained the characteristics of the individual detergents, depending on their concentrations.
BradfordMM . 1976 . Rapid and Sensitive Method for Quantitation of Microgram Quantities of Protein Utilizing Principle of Protein-dye Binding . Anal Biochem , 72 : 248 - 254 .
BrewisIA , GadellaBM . 2010 . Sperm surface proteomics: from protein lists to biological function . Mol Hum Reprod , 16 : 68 - 79 .
D’AmoursO , FrenetteG , FortierM , LeclercP , SullivanR.2010 . Proteomic comparison of detergent-extracted sperm proteins from bulls with different fertility indexes . Reproduction , 139 : 545 - 556 .
Du PlessisSS , KashouAH , BenjaminDJ , YadaSP , AgarwalA . 2011 . Proteomics: a subcellular look at spermatozoa . Reprod Biol Endocrinol , 9 : 36 - 47 .
GingrasAG , GstaigerM , RaughtB , AebersoldR . 2007 . Analysis of protein complexes using mass spectrometry . Nature Reviews Molecular Cell Biology , 8 : 645 - 654 .
GirouardJ , FrenetteG , SullivanR.2009 . Compartmentalization of Proteins in Epididymosomes Coordinates the Association of Epididymal Proteins with the Different Functional Structures of Bovine Spermatozoa . Biol Reprod , 80 : 965 - 972 .
GuoW , QuF , XiaL , GuoQ , YingX , DingZ.2007 . Identification and characterization of ERp29 in rat spermatozoa during epididymal transit . Reproduction , 133 : 575 - 584 .
HochstrasserDF , HarringtonMG , HochstrasserAC , MillerMJ , MerrilCR . 1988 . Methods for Increasing the Resolution of Two-Dimensional Protein Electrophoresis . Anal Biochem , 173 : 424 - 435 .
IgnotzGG , LoMC , PerezCL , GwathmeyTM , SuarezSS . 2001 . Characterization of a Fucose-Binding Protein from Bull Sperm and Seminal Plasma That May Be Responsible for Formation of the Oviductal Sperm Reservoir . Biol Reprod , 64 : 1806 - 1811 .
IjiriTW , MerdiushevT , CaoW , GertonGL . 2011 . Identification and validation of mouse sperm proteins correlated with epididymal maturation . Proteomics , 11 : 4047 - 4062 .
JakopU , FuchsB , SubR , WibbeltG , BraunB , MullerK , SchillerJ.2009 . The solubilisation of boar sperm membranes by different detergents - a microscopic, MALDI-TOF MS, 31P NMR and PAGE study on membrane lysis, extraction efficiency, lipid and protein composition . Lipids Health Dis , 8 : 49 - 65 .
PaaschU , HeidenreichF , PurscheT , KuhlischE , KettnerK , GrunewaldS , KratzschJ , DittmarG , GlanderHJ , HoflackB , KriegelTM . 2011 . Identification of Increased Amounts of Eppin Protein Complex Components in Sperm Cells of Diabetic and Obese Individuals by Difference Gel Electrophoresis . Mol Cell Proteomics , 10 : M110.007187 .
PeddintiD , NanduriB , KayaA , FeugangJM , BurgessSC , MemiliE.2008 . Comprehensive proteomic analysis of bovine spermatozoa of varying fertility rates and identification of biomarkers associated with fertility . BMC Syst Biol , 2 : 19 - 31 .
PerkinsDN , PappinDJC , CreasyDM , CottrellJS . 1999 . Probability-based protein idenification by searching sequence databases using mass spectrometry data . Electrophoresis , 20 : 3551 - 3567 .
RajeevSK , ReddyKVR . 2004 . Sperm membrane protein profiles of fertile and infertile men: identification and characterization of fertility-associated sperm antigen . Hum Reprod , 19 : 234 - 242 .
ReeseKL , AravindanRG , Griffiths , GS , ShaoM , WangY , GalileoDS , AtmuriV , Triggs-RaineBL , Martin-DeleonPA . 2010 . Acidic Hyaluronidase Activity Is Present in Mouse Sperm and Is Reduced in the Absence of SPAM1: Evidence for a Role for Hyaluronidase 3 in Mouse and Human Sperm . Mol Reprod Dev , 77 : 759 - 772 .
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