scholarly journals Sperm Proteomics: Road to Male Fertility and Contraception

2013 ◽  
Vol 2013 ◽  
pp. 1-11 ◽  
Author(s):  
Md Saidur Rahman ◽  
June-Sub Lee ◽  
Woo-Sung Kwon ◽  
Myung-Geol Pang
Keyword(s):  
Posttranslational Modifications ◽  
Gel Electrophoresis ◽  
Male Fertility ◽  
Large Scale ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein Biomarkers ◽  
Scientific Publications ◽  
Pubmed Database ◽  
Protein Functions ◽  
Specific Proteins

Spermatozoa are highly specialized cells that can be easily obtained and purified. Mature spermatozoa are transcriptionally and translationally inactive and incapable of protein synthesis. In addition, spermatozoa contain relatively higher amounts of membrane proteins compared to other cells; therefore, they are very suitable for proteomic studies. Recently, the application of proteomic approaches such as the two-dimensional polyacrylamide gel electrophoresis, mass spectrometry, and differential in-gel electrophoresis has identified several sperm-specific proteins. These findings have provided a further understanding of protein functions involved in different sperm processes as well as of the differentiation of normal state from an abnormal one. In addition, studies on the sperm proteome have demonstrated the importance of spermatozoal posttranslational modifications and their ability to induce physiological changes responsible for fertilization. Large-scale proteomic studies to identify hundreds to thousands of sperm proteins will ultimately result in the development of novel biomarkers that may help to detect fertility, the state of complete contraception, and beyond. Eventually, these protein biomarkers will allow for a better diagnosis of sperm dysfunctions and aid in drug development. This paper reviews the recent scientific publications available from the PubMed database to address sperm proteomics and its potential application to characterize male fertility and contraception.

scholarly journals Deciphering and engineering chromodomain-methyllysine peptide recognition

2018 ◽  
Vol 4 (11) ◽  
pp. eaau1447 ◽  
Author(s):  
Ryan Hard ◽  
Nan Li ◽  
Wei He ◽  
Brian Ross ◽  
Gary C. H. Mo ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Posttranslational Modifications ◽  
Large Scale ◽  
Regulation Of Gene Expression ◽  
Live Cells ◽  
Lysine Methylation ◽  
Dynamic Regulation ◽  
Peptide Recognition ◽  
Protein Functions ◽  
Domain Peptide

Posttranslational modifications (PTMs) play critical roles in regulating protein functions and mediating protein-protein interactions. An important PTM is lysine methylation that orchestrates chromatin modifications and regulates functions of non-histone proteins. Methyllysine peptides are bound by modular domains, of which chromodomains are representative. Here, we conducted the first large-scale study of chromodomains in the human proteome interacting with both histone and non-histone methyllysine peptides. We observed significant degenerate binding between chromodomains and histone peptides, i.e., different histone sites can be recognized by the same set of chromodomains, and different chromodomains can share similar binding profiles to individual histone sites. Such degenerate binding is not dictated by amino acid sequence or PTM motif but rather rooted in the physiochemical properties defined by the PTMs on the histone peptides. This molecular mechanism is confirmed by the accurate prediction of the binding specificity using a computational model that captures the structural and energetic patterns of the domain-peptide interaction. To further illustrate the power and accuracy of our model, we used it to effectively engineer an exceptionally strong H3K9me3-binding chromodomain and to label H3K9me3 in live cells. This study presents a systematic approach to deciphering domain-peptide recognition and reveals a general principle by which histone modifications are interpreted by reader proteins, leading to dynamic regulation of gene expression and other biological processes.

scholarly journals Identification of α1-Antitrypsin Variants in Plasma with the Use of Proteomic Technology

2001 ◽  
Vol 47 (11) ◽  
pp. 2012-2022 ◽  
Author(s):  
Kevin Mills ◽  
Philippa B Mills ◽  
Peter T Clayton ◽  
Andrew W Johnson ◽  
David B Whitehouse ◽  
...  
Keyword(s):  
Posttranslational Modifications ◽  
Metabolic Diseases ◽  
Polyacrylamide Gel Electrophoresis ◽  
Complete Characterization ◽  
Proteomic Technology ◽  
Flight Mass Spectrometry ◽  
Type Ia ◽  
Specific Proteins ◽  
Polypeptide Structure

Abstract Background: Proteomic technology permits the investigation of genetic metabolic diseases at the level of protein expression. Changes in the expression, polypeptide structure, and posttranslational modification of individual proteins can be detected in complex mixtures of proteins. Methods: We used high-resolution two-dimensional polyacrylamide gel electrophoresis to separate isoforms of plasma proteins and detect abnormalities of mass and/or charge. We confirmed the identity of the separated proteins by in-gel digestion with proteases and N-glycanases and then analyzed the released peptides and glycans by matrix-assisted laser-desorption ionization–time-of-flight mass spectrometry. Results: Complete characterization of the polypeptide sequences and glycosylation of α1-antitrypsin isoforms was achieved in plasma from controls and from patients with three different known α1-antitrypsin deficiencies and congenital disorder of glycosylation type Ia. Conclusions: This study shows that proteomic techniques are a powerful and sensitive means of detecting changes in the amino acid sequence and abnormal posttranslational modifications of specific proteins in a complex biologic matrix.

scholarly journals Clustering of Clinical Strains ofHelicobacter pylori Analyzed by Two-Dimensional Gel Electrophoresis

2000 ◽  
Vol 7 (2) ◽  
pp. 301-306 ◽  
Author(s):  
Helena Enroth ◽  
Thomas Åkerlund ◽  
Anna Sillén ◽  
Lars Engstrand
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Two Dimensional ◽  
Strain Variation ◽  
Disease Group ◽  
Two Dimensional Gel Electrophoresis ◽  
Specific Proteins ◽  
Patient Groups ◽  
H Pylori ◽  
Disease Specific

ABSTRACT Strain variations of Helicobacter pylori have been tested by numerous methods and compared among different patient groups. The aim of this study was to investigate whether H. pyloriexpresses disease-specific proteins that can be detected by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). H. pylori strains isolated from duodenal ulcer, gastric cancer, and gastritis patients were analyzed. Extensive variation in spot patterns was observed between the strains, but a dendrogram analysis revealed that some strains within each disease group clustered together. Eight proteins were sequenced and found in the H. pylori genome sequence. 2-D PAGE is a useful method for studies of protein expression and for highlighting the extensive strain variation that H. pylori exhibits.

scholarly journals Calcium Influx and Male Fertility in the Context of the Sperm Proteome: An Update

2014 ◽  
Vol 2014 ◽  
pp. 1-13 ◽  
Author(s):  
Md Saidur Rahman ◽  
Woo-Sung Kwon ◽  
Myung-Geol Pang
Keyword(s):  
Male Fertility ◽  
Calcium Influx ◽  
Protein Protein Interaction ◽  
Channel Proteins ◽  
Sperm Proteins ◽  
Pubmed Database ◽  
Protein Functions ◽  
Ion Channel Proteins ◽  
Insight Into ◽  
Mammalian Spermatozoa

Freshly ejaculated spermatozoa are incapable or poorly capable of fertilizing an oocyte. The fertilization aptness of spermatozoa depends on the appropriate and time-dependent acquisition of hyperactivation, chemotaxis, capacitation, and the acrosome reaction, where calcium (Ca2+) is extensively involved in almost every step. A literature review showed that several ion channel proteins are likely responsible for regulation of the Ca2+uptake in spermatozoa. Therefore, manipulation of the functions of channel proteins is closely related to Ca2+influx, ultimately affecting male fertility. Recently, it has been shown that, together with different physiological stimuli, protein-protein interaction also modifies the Ca2+influx mechanism in spermatozoa. Modern proteomic analyses have identified several sperm proteins, and, therefore, these findings might provide further insight into understanding the Ca2+influx, protein functions, and regulation of fertility. The objective of this review was to synthesize the published findings on the Ca2+influx mechanism in mammalian spermatozoa and its implications for the regulation of male fertility in the context of sperm proteins. Finally, Pathway Studio (9.0) was used to catalog the sperm proteins that regulate the Ca2+influx signaling by using the information available from the PubMed database following a MedScan Reader (5.0) search.

scholarly journals Rat hepatic microsomal cytochrome b5. A simple large-scale purification procedure and antibody production by antigen-containing liposomes

1988 ◽  
Vol 256 (3) ◽  
pp. 1051-1054 ◽  
Author(s):  
J Carlsen ◽  
K Christiansen ◽  
H M Jensen
Keyword(s):  
Gel Electrophoresis ◽  
Large Scale ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cytochrome B5 ◽  
Liver Microsomes ◽  
Lipid Vesicles ◽  
Rat Liver Microsomes ◽  
Native Form ◽  
Great Capacity ◽  
Microsomal Cytochrome B5

Cytochrome b5 from rat liver microsomes (microsomal fractions) was purified in its native form. The procedure described has great capacity, is fast, and the final product is pure as judged from SDS/polyacrylamide-gel electrophoresis. Antibodies to cytochrome b5 are obtained after administration of the antigen inserted into small unilamellar lipid vesicles.

scholarly journals Proteomic Characterisation of Lupin (Lupinus angustifolius) Milk as Influenced by Extraction Techniques, Seed Coat and Cultivars

Molecules ◽  
2020 ◽  
Vol 25 (8) ◽  
pp. 1782
Author(s):  
Nadia Al-Saedi ◽  
Manjree Agarwal ◽  
Wujun Ma ◽  
Shahidul Islam ◽  
Yonglin Ren
Keyword(s):  
Gel Electrophoresis ◽  
Milk Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Two Dimensional ◽  
Extraction Techniques ◽  
Centrifuge Method ◽  
Lupin Seeds ◽  
Specific Proteins ◽  
Protein Extractability ◽  
Better Than

Lupin seeds are rich in proteins and other essential ingredients that can help to improve human health. The protein contents in both whole and split seeds of two lupin cultivars (Mandleup and PBA Jurien) were used to produce the lupin milk using the cheesecloth and centrifuge method. Proteins were extracted from the lupin milk using thiourea/urea solubilization. The proteins were separated by a two-dimensional polyacrylamide gel electrophoresis and then identified with mass spectrometry. A total of 230 protein spots were identified, 60 of which showed differential abundances. The cheesecloth separation showed protein extractability much better than that of the centrifuge method for both the cultivars. The results from this study could offer guidance for future comparative analysis and identification of lupin milk protein and provide effective separation technique to determine specific proteins in the cheese-making process.

Protein Synthesis In Megakaryocytes Stimulated By Thrombo- Poietin In Vitro

1981 ◽  
Author(s):  
K L Kellar ◽  
B L Evatt ◽  
C R McGrath ◽  
R B Ramsey
Keyword(s):  
Protein Synthesis ◽  
Guinea Pig ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Leucine Incorporation ◽  
Ploidy Levels ◽  
Percoll Gradients ◽  
Specific Proteins ◽  
Phosphate Buffered Saline

Studies in our laboratories have been concerned with the responses of megakaryocytes to thrombopoietin in vitro. We have shown that preparations of thrombopoietin stimulate DNA synthesis in guinea pig megakaryocytes. The increase in 3H-thymidihe incorporation correlates with an increase in the labeling index of the megakaryocytes. After 2 and 3 days of incubation an increase in the ploidy levels of the megakaryocytes has been observed in thrombo- poietin-supplemented cultures compared to controls.Recent studies have examined the incorporation of 3H-leucine in megakaryocyte cultures. Megakaryocytes were prepared on BSA or Percoll gradients to purities of 70-95%. 3H-leucine incorporation was measured after a 15 hr incubation of the megakaryocytes in medium containing 10% thrombopoietin or control preparations of normal plasma or phosphate-buffered saline. Utilization of isotope increased over a 24 hr period and was higher in the thrombopoietin-supplemented cultures. In addition, synthesis of specific proteins was analyzed by using SDS- polyacrylamide gel electrophoresis and quantitation was achieved by employing rocket immunoelectrophoresis. The results indicate that thrombopoietin stimulates endoredu- plication and protein synthesis in megakaryocytes in vitro and that this system may serve as a model for studying the mechanism of action of thrombopoietin in megakaryocytopoiesis.

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