Relationship between seminal plasma tuberoinfundibular peptide of 39 residues and sperm functional attributes in buffalo (Bubalus bubalis)

2016 ◽  
Vol 28 (10) ◽  
pp. 1622 ◽  
Author(s):  
Sellappan Selvaraju ◽  
Lakshminarayana Somashekar ◽  
Binsila B. Krishnan ◽  
Sivashanmugam Parthipan ◽  
Guvvala Pushparani ◽  
...  
Keyword(s):  
Plasma Protein ◽  
Seminal Plasma ◽  
Sperm Quality ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Profiling ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Integrity Test ◽  
Tuberoinfundibular Peptide

The buffalo seminal plasma protein profile and its relationship with sperm quality have not been studied in detail. Thus, the aim of the present study was to profile buffalo seminal plasma proteins and to assess the relationship between differentially expressed proteins and sperm characteristics. Semen samples (n = 44) were collected from 11 Murrah buffalo bulls (four ejaculates from each animal) and seminal plasma protein profiling was performed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Matrix-assisted laser desorption ionisation time-of-flight analysis of one of the differentially expressed proteins, namely the 11–12 kDa protein, identified it as tuberoinfundibular peptide of 39 residues (TIP39). Western blot analysis confirmed the presence of TIP39, with TIP39 expression in seminal plasma varying among bulls. Based on TIP39 levels, bulls were classified into two groups, those with high and low protein. The percentages of spermatozoa positive for mitochondrial membrane potential test, chromatin distribution test, synthetic media sperm penetrability test and acrosomal integrity test were significantly (P < 0.05) high in the high protein group. The present study is the first to demonstrate the presence of TIP39 in buffalo seminal plasma and the positive effect of TIP39 on the functional parameters and fertilising ability of spermatozoa.

Comparative Analysis of Soluble Proteins in Four Medicinal Aloe Species by Two-Dimensional Electrophoresis and MALDI-TOF-MS

2019 ◽  
Vol 102 (3) ◽  
pp. 748-760 ◽  
Author(s):  
Jiao-Jiao Fan ◽  
Chun-Hong Li ◽  
Guang Hu ◽  
Cheng-Ning Tan ◽  
Feng-Qing Yang ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Protein Extraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Morphological Characteristics ◽  
Soluble Proteins ◽  
Differentially Expressed ◽  
Two Dimensional ◽  
Differentially Expressed Proteins ◽  
Phenol Extraction

Abstract Background: Aloe barbadensis Miller 1768, A. vera L. var. chinensis (Haw.) Berger 1908, A. ferox Miller 1768, and A. arborescens Miller 1768 are the most widely cultivated species of Aloe and are used in Asia along with 400 other Aloe species worldwide because of their potent and potential bioactivity. Objective: The objective was to analyze and compare the soluble proteins of four commonly used medicinal Aloe species. Methods: Aloe protein samples were obtained by TCA/acetone-saturated phenol-methanol/ammonium acetate combined extraction (phenol extraction), and then were analyzed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and two-dimensional gel electrophoresis. Finally, the differentially expressed proteins of four Aloe species were identified by matrix-assisted laser desorption ionization–time-of-flight–MS analysis. Results: The phenol extraction method was the most suitable method for the protein extraction of Aloe. Fifty differentially expressed proteins in four Aloe species were successfully identified and divided into eight functional categories. Furthermore, Malate dehydrogenase and ran-binding protein in A. barbadensis, cytoskeletal-related protein tubulin in A. vera var. chinensis and auxin-induced protein PCNT-115 in A. arborescens are closely related to their morphological characteristics. Conclusions: There are differences in the soluble proteins of the four Aloe species. Those proteins, related to the difference of their morphology of Aloe, might be used to identify different species. Highlights: Fifty differentially expressed proteins in four medicinal Aloe species were identified, and these proteins were classified into eight categories according to their biological functions. Four special proteins closely related to the morphological characteristics of Aloe were found and might be used to identify these four Aloe species.

scholarly journals Evaluation of effective protein extraction procedure to profile petiole of Moringa oleifera

2020 ◽  
Vol 7 (2) ◽  
pp. 214
Author(s):  
Zetty Amirah Zulkifli ◽  
Zaidah Rahmat
Keyword(s):  
Moringa Oleifera ◽  
Protein Extraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Extraction Procedure ◽  
Electrophoretic Pattern ◽  
Antioxidant Property ◽  
Extraction Methods ◽  
Protein Profiling ◽  
Sds Page

Moringa oleifera is widely known as multipurpose tree since all of its parts confer multiple functions. The leaf is highly favourable among consumers while the petiole is mostly wasted. There are numerous studies on the flavonoid and antioxidant property of the stem and twig. However, study on the petiole has never been done. There-upon, this study was conducted to develop protein profiling of the petiole. In this study, 6 different protein extraction methods were tested on the fresh petiole before its protein quantity and quality were checked via Bradford assay and Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) respectively. The in-solution digestion was then done prior to LC-MS/MS analysis. The protein electrophoretic pattern from the SDS-PAGE proves that method 6 using Tris HCl buffer with incorporation of dithiothreitol (DTT) and phenylmethylsulfonyl fluoride (PMSF) confers the best quality of protein. It produced the highest number of visible individual bands compared to other methods. Meanwhile, 93 proteins were successfully identified via LCMS analysis where the protein, signal response and carbohydrate metabolism categories confer the highest percentage. High quality and content of the protein extracted from the petiole including the antioxidant, anticancer and antidiabetic protein identified suggested that consuming this part of the plant could enhance nutrients of human body.

Heparin Binding Proteins of Black Bengal Buck Semen and Their Correlation with Sperm Characters and Freezability

2019 ◽  
Author(s):  
M Karunakaran ◽  
Vivek C Gajare ◽  
Ajoy Mandal ◽  
Mohan Mondal ◽  
S K Das ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Binding Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Heparin Binding ◽  
Sds Page ◽  
Significant Difference ◽  
Sperm Characters

This experiment was conducted to study the electrophoretic characters of heparin binding proteins (HBP) of Black Bengal buck semen and their correlation with sperm characters and cryo-survivability. Semen ejaculates (n=20/buck) were collected from nine bucks and in vitro sperm characters were evaluated at collection, after equilibration and after freeze - thawing. HBP were isolated through heparin column and discontinuous Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) was performed to assess molecular weight. Significant difference (plessthan0.01) were observed among the bucks in sperm characters and freezability. Eight protein bands of 17 to 180 kDa in seminal plasma and 7 bands in sperm were found. 180 -136 kDa HBP of seminal plasma and 134-101 kDa HBP of sperm had showed high correlation with in vitro sperm characters. Further studies on identification of these proteins and their correlation with in vivo pregnancy are needed to find their role as marker for buck selection.

scholarly journals Heterogeneity of Mycoplasma Iowae Determined by Restriction Enzyme Analysis

1989 ◽  
Vol 1 (2) ◽  
pp. 165-169 ◽  
Author(s):  
Shaohua Zhao ◽  
Richard Yamamoto
Keyword(s):  
High Titer ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Inhibition Test ◽  
Protein Profiling ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Protein Patterns ◽  
Biochemical Tests ◽  
Mycoplasma Iowae

Strains of Mycoplasma iowae were homogeneous in some characteristics and heterogeneous in others. Thus, the biochemical tests, immunofluorescence, and protein profiling by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were group-specific tests. However, some minor differences in protein patterns were seen among strains. The growth inhibition test tended to be strain-specific. Hemagglutination titers were very low and unstable for the majority of strains. One strain (RY-65) with a stable high-titer hemagglutinin failed to react in the hemagglutination-inhibition test against immune sera to the reference strains. Restriction endonuclease DNA analyses was the most useful method to differentiate 1 strain from another.

HSP90 expression correlation with the freezing resistance of bull sperm

Zygote ◽  
2013 ◽  
Vol 22 (2) ◽  
pp. 239-245 ◽  
Author(s):  
Peng Wang ◽  
Yan-Feng Wang ◽  
Hong Wang ◽  
Chun-Wei Wang ◽  
Lin-Sen Zan ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Freezing Resistance ◽  
Hsp90 Expression ◽  
Sds Page ◽  
Associated Proteins ◽  
Holstein Bulls ◽  
Bull Sperm

SummaryTo date, there has been little improvement in cryopreservation of bull sperm due to lack of understanding of the freezing mechanisms. Therefore, this study set out to investigate expression levels of fertility-associated proteins in bull sperm, and in particular the relationship between the 90 kDa heat-shock protein (HSP90) and the sperm characteristics after freezing–thawing. Semen was collected from eight Holstein bulls by artificial vagina. Characteristics of these fresh semen, including sperm motility, morphology, viability and concentration, were evaluated. Sperm quality was also assessed after freezing–thawing. Eight ejaculates were divided into two groups based on freezing resistance and sperm motility. Sperm proteins were extracted and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis and western blotting were performed. SDS-PAGE results showed that there was substantial diversity in 90 kDa proteins in the frozen–thawed sperm and HSP90 was confirmed as one of the 90 kDa proteins by western blot. This study indicated that HSP90 expression correlated positively with sperm quality. The amount of expressed 90 kDa proteins in the high freezing resistance (HFR) group was significantly higher than that in the low freezing resistance (LFR) group (P< 0.05). Thus, higher expression of HSP90 could probably lead to the higher motility and freezing resistance of sperm found after freezing–thawing. Therefore, we concluded that level of HSP90 expression could be used to predict reliably and simply the freezing resistance of bull sperm.

scholarly journals A Comparative Study of Distribution of Protein and Cholesterol in Various Fractions of Human Semen from Infertile and Fertile Subjects

2012 ◽  
Vol 3 (3) ◽  
pp. 78-82 ◽  
Author(s):  
MS Srinivas ◽  
Vickram Sundaram ◽  
M Ramesh Pathy ◽  
TB Sridharan
Keyword(s):  
Molecular Weight ◽  
Comparative Study ◽  
Seminal Plasma ◽  
Semen Analysis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Human Semen ◽  
Exact Function ◽  
Wide Range ◽  
Significant Difference

ABSTRACT Aim To elucidate the concentration of the protein and cholesterol in different fractions of human semen from different infertile categories and comparing them with the fertile group. Materials and methods The human semen was collected from different infertile categories including oligoasthenospermia, asthenospermia, azoospermia, normospermia, oligospermia and fertile group. Immediately after collection, the semen analysis was done as per WHO standard protocols. After that, the semen was centrifuged to get the different fractions. Four main fractions were obtained, (1) spermatozoa, (2) debris or material that precipitates at 12 K rpm for 10 minutes, (3) prostasomes which was precipitated at 20K rpm for 120 minutes, (4) seminal plasma. The protein concentration was done by Lowry's method and cholesterol was estimated by diagnostic kit. Results Sodium dodecyl sulfate—polyacrylamide gel electrophoresis (SDS PAGE) was run for all the categories of semen samples for their seminal plasma and the fertility associated protein was identified. A significant difference was found in the concentration of proteins in all subfractions when compared between control and infertile categories. Almost 86% of the protein was recovered from soluble fraction. In case of azoospermia, the protein content was very low when compared with fertile group. Seminal plasma proteins were visualized by silver staining. The molecular weight of the protein bands were ranging from 6.5 to 205 kDa. The band with molecular weight around 55 kDa was found to be missing in case of oligoasthenospermia. This particular protein is said to be fertility associated protein. The content of cholesterol for different subfraction of the human semen samples from infertile and fertile samples was compared. A wide range of cholesterol was recovered from prostasomes, that too purified. Conclusion A thrive study have to be done in all the subfractions of the semen irrespective of the category of samples to know the exact function of the each subfractions in terms of protein and cholesterol distribution. How to cite this article Sundaram V, Srinivas MS, Rao KA, Pathy MR, Sridharan TB. A Comparative Study of Distribution of Protein and Cholesterol in Various Fractions of Human Semen from Infertile and Fertile Subjects. Int J Infertility Fetal Med 2012;3(3):78-82.

scholarly journals Identification of Differentially Expressed Protein from Electrical Stunning of Broiler Chickens Meat Protein

Molekul ◽  
2016 ◽  
Vol 11 (1) ◽  
pp. 71 ◽  
Author(s):  
Sandra Hermanto ◽  
Maya Ina Sholaikah ◽  
Sri Suci Mulyani
Keyword(s):  
Muscle Tissue ◽  
Broiler Chickens ◽  
Broiler Chicken ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Specific Protein ◽  
Differentially Expressed ◽  
Over Expression ◽  
Differentially Expressed Protein ◽  
Sds Page

Identification of differentially expressed protein from the muscle tissue of broiler chicken meat with different conditions of pre-slaughter has been done. Each sample (6 broilers aged 21 days, 1 kg of weight ) was prepared through the process of pre-slaughter with 3 conditions, the first sample slaughtered in a conventional way which untreated electrical stunning, while the second and third sample of the chicken was prepared by using electrical stunning with 1 A and 25 Volts for 5 seconds and 1 A, 125 Volts for 30 seconds. Two biological replicate were done for each of samples. Muscle tissue protein extracted in Tris HCl pH 8.0 and the proteins separation by using SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis). Identification of differentially expressed protein performed by densitometry to identify the profile of the resulting proteins. The results of this study showed that the protein bands constructed in the range of 8.5-140 kDa and 9 dominant protein bands with different relative intensities. Densitogram analysis results showed there are two specific protein bands appear on the results of the electrical stunning which more extensive over expression. This indicates the electrical stunning of slaughter process may triggered the expression levels of certain proteins that do not occur in the nonelectrical stunning.

COMPARISON OF ALLERGENIC PROTEINS OF SEA SNAIL (CERITHIDEA OBTUSA) AND FRESHWATER SNAIL (POMACEA CANALICULATA)

2016 ◽  
Vol 78 (11) ◽  
Author(s):  
Rosmilah Misnan ◽  
Norazlin Salahudin Abdul Aziz ◽  
Zailatul Hani Mohd Yadzir ◽  
Noormalin Abdullah ◽  
Faizal Bakhtiar ◽  
...  
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Freshwater Snail ◽  
Protein Profiling ◽  
Protein Fractions ◽  
Pomacea Canaliculata ◽  
Market Demand ◽  
Allergenic Proteins ◽  
Major Allergens ◽  
Minor Allergens

In Malaysian and certain Asian countries, snail has high market demand and popular to the local people as food. However, snail is also frequently reported as one of the worst food allergens, dominated by severe symptoms such as asthma and anaphylactic shock. Thus, the aims of this study is to determine the allergenicity of two species of edible snails; the local sea snail, Cerithidea obtusa and the freshwater snail Pomacea canaliculata. Snail extracts were prepared from the snail flesh and analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to determine their protein profiling. Allergenic proteins were detected by immunoblotting test using sera from 10 snail-allergic patients. The snails contain 31 to 34 protein fractions between 11 to >250 kDa. The prominent bands were seen at 33, 42, 74 and 250 kDa. Immunoblotting detected 15 and 16 allergenic proteins in C. obtusa and P. canaliculata, respectively. Three protein fractions at 30, 33 and 42 kDa were identified as the major allergens of C. obtusa, while six major allergens at 30, 33, 42, 74, 124 and 218 kDa were detected in P. canaliculata. Various minor allergens were also detected in both snails. This study indicated that numerous proteins of C. obtusa and P. canaliculata were allergenic. Thus, combined allergen extracts of both snails are essential to be included in diagnosis of snail allergy among local allergic patients.  

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