Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P56851 (epididymal)
 11,273 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The triazine dyes, Cibacron blue F3GA and Procion red HE3B inhibited diaphorase activity of ferredoxin-NADP+ reductase, in a competitive manner with respect to NADPH. The Ki values were 1.5 and 0.2 microM, respectively. Binding of the dyes to the flavoprotein, as measured by difference spectroscopy, indicated an apparent stoichiometry of 1 mol dye/mol reductase and was prevented by NADP+ or high ionic strength. Chemical modification of a lysine residue and a carboxyl group at the NADP(H) binding site of the enzyme prevented complex formation with Procion red. Procion red showed a higher affinity for ferredoxin-NADP+ reductase than Cibacron blue. The Kd values were 1.9 and 5 microM, respectively. Once covalently linked to a Sepharose matrix, the triazine compounds specifically bind the flavoprotein. The interaction is partially electrostatic and partially hydrophobic. The enzyme can be eluted by high concentrations of salt or low concentrations of the corresponding coenzyme. The use of this affinity column allows the rapid purification of ferredoxin-NADP+ oxidoreductase from spinach leaves with good yields.
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PMID:Interaction of ferredoxin-NADP+ oxidoreductase with triazine dyes. A rapid purification method by affinity chromatography. 682 90

A new melanocyte-stimulating peptide has been isolated from acid extracts of frozen human pituitary glands by salt/ethanol fractionation, Sephadex G-75 gel filtration and DEAE- and cM-cellulose ion-exchange chromatography. The peptide is glycosylated, has an N-terminal tryptophan residue and an apparent mol.wt. of 16000 as estimated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Its amino acid analysis closely resembles residues Trp-105 to Gln-29 predicted for the common precursor protein of bovine corticotropin and beta-lipotropin by Nakanishi, Inoue, Kita, Nakamura, Chang, Cohen & Numa [(1979) Nature (London) 278, 423-427]. This fragment is expected to have melanotropin activity due to the tetrapeptide -His-Phe-Arg-Trp- (residues -51 to -48) of the predicted sequence of the common precursor. It was found to have a molar potency of 1 X 10(-5) relative to alpha-melanotropin in the frog skin bioassay. These characteristics are consistent with the isolated melanotropin peptide being a non-corticotropin, non-lipotropin peptide of the human common precursor protein of corticotropin and lipotropin. The peptide neither potentiates the adrenal weight-maintenance activity of corticotropin-(1-24)-tetracosapeptide when administered to hypophysectomized rats, nor stimulates release of non-esterified fatty acids from isolated rat epididymal cells. A second N-terminal-tryptophan glycopeptide was also isolated, which had an amino-acid composition similar to that predicted for the bovine common precursor protein, residues Trp-105 to Gly-35.
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PMID:Purification and characterization of a gamma-melanotropin precursor from frozen human pituitary glands. 747 89

The localization of Trimetaphosphatase (TMPase) activity during the acrosomal formation in the mouse testis was enzyme cytochemically investigated by the cerium-salt method. In addition to the lysosomes of the Sertoli cells and the spermatogenic cells in the seminiferous tubules, positive TMPase activity was detected in the Golgi complex and in the acrosomal vesicles of the spermatids, as well as in the acrosomes of both spermatids and spermatozoa. In the Golgi complex of the spermatids, TMPase activity was observed in the first one or two lamellae of the trans-face and in the small vesicles in the vicinity of the Golgi complex. TMPase positive reaction was also detected in the acrosomes of the spermatozoa in the lumina of both the seminiferous tubules and the epididymal duct. The localization of this enzyme activity was compared with that of acid phosphatase (ACPase), as detected by the cerium-based method, using beta-glycerophosphate as substrate: ACPase activity was completely absent from the Golgi complex, small vesicles, acrosomal vesicle and acrosome throughout the entire process of acrosomal formation. TMPase is thought to become one of the acrosomal components, and may be involved in the acrosomal reaction during fertilization.
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PMID:Ultracytochemical study of trimetaphosphatase activity during acrosomal formation in the mouse testis. 757 18

A protein kinase that causes phosphorylation of serine and threonine residues of casein has been partially purified from goat cauda-epididymal sperm plasma membrane and characterized. The kinase, solubilized from the membrane with 1.0% Triton X-100, was purified to 480-fold by using DEAE-cellulose and casein-Sepharose affinity chromatographic techniques. The kinase is a strongly basic protein with pI of 9.5. The enzyme has a molecular mass of 310 kilodaltons as estimated by Sephacryl S-300 gel exclusion. The kinase showed affinity for protein substrates in the order membrane proteins > casein > phosvitin > histone > protamine. The apparent Km values of the kinase for casein and membrane proteins were 1 and 0.15 mg/mL, respectively. The synthetic peptides Kemptide and poly(Glu80Tyr20) did not serve as substrates of the enzyme. ATP, rather than GTP or PP(i), is the donor of phosphate for the phosphorylation reaction. Cyclic AMP and GMP, NaCl (0.25 M), KCl (0.25 M), Ca2+, calmodulin, phosphatidylserine, and muscle protein kinase inhibitor had no appreciable effect on the kinase activity. Heparin (0.5 microgram/mL) showed high affinity for inhibiting only 40% of the kinase activity, whereas polyamines at a relatively high concentration (5 mM) inhibited 40-50% of the enzymic activity. The kinase appears to be distinct from other protein kinases including casein kinases. The activity of the kinase derived from the purified sperm plasma membrane was markedly (approximately 90%) lost when the intact spermatozoa were pretreated with diazonium salt of sulfanilic acid, a membrane nonpenetrating surface probe.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Purification and characterization of a protein kinase from goat sperm plasma membrane. 784 Sep 41

The cellular origin of a soluble phospholipid-binding protein (PBP) in rat testicular and epididymal secretions has been investigated genetically and immunologically. PBP is ubiquitous in tissue cytosols but is not present in blood serum, lymph or milk. The relatively large amounts present in cauda epididymal plasma (CEP) and rete testis fluid suggested therefore that it may be secreted specifically by these tissues. However, when PBP cDNAs from testis and epididymis were cloned and sequenced, they did not contain a signal peptide and only one size of transcript was obtained on Northern blots of RNAs from liver, brain, placenta, testis and epididymis. Moreover, PBP could not be detected in sperm-free CEP from castrated, androgen-stimulated animals or in medium from Sertoli cell cultures. Spermatozoa, on the other hand, contained significant amounts of PBP that could be solubilized by washing cells in dissociating reagents or high-salt solutions. These results indicate that, contrary to previous interpretations, PBP is not secreted by classical pathways in either the testis or epididymis but that its presence in CEP and rete testis fluid is attributable largely to release from spermatozoa. Thus, spermatozoa have a significant influence on the composition of CEP as well as on the secretory and absorptive activity of the epididymal epithelium. A possible role for PBP in membrane biogenesis and maintenance of antigen segregation in spermatozoa is discussed.
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PMID:Sequence analysis of a mammalian phospholipid-binding protein from testis and epididymis and its distribution between spermatozoa and extracellular secretions. 803 77

Based upon recent reports that the mRNA from the regulatory (R) RI beta subunit of cAMP-dependent protein kinase (PKA) was expressed in testicular extracts, we determined whether testicular extracts exhibited RI beta protein. To accomplish this goal, we initially determined the fundamental labeling and ionic characteristics of recombinant RI beta. Recombinant RI beta eluted from DEAE-cellulose with a salt concentration (of 0.075 M) equivalent to its elution position from soluble mouse brain extracts with catalytic subunit-free RI alpha. As predicted by its amino acid sequence homology to RI alpha, recombinant RI beta was not phosphorylated by PKA but was labeled specifically with 8-azido-adenosine 3':5'-[32P]monophosphate (8-N3[32P]cAMP). Additionally, RI antisera reacted equally with RI alpha (47 kDa) and recombinant RI beta (53 kDa). However, recombinant RI beta exhibited an unexpectedly basic pI of 6.65-6.85. By using a pH gradient for isoelectric focussing that allowed for clear focussing of 8-N3[32P]cAMP-labeled recombinant RI beta, 8-N3[32P]cAMP-labeled RI beta was readily detected by two-dimensional gel electrophoresis in rat brain particulate extracts and exhibited a pI equivalent to that of recombinant RI beta. The 53-kDa RI beta was undetectable either by its immunoreactivity or upon photoaffinity labeling with 8-N3[32P]cAMP by one or two-dimensional gel electrophoresis in soluble or particulate extracts of testes of 14-day-old, 45-day-old, or adult rats or in epididymal sperm. However, 8-N3[32P]cAMP-labeled RI beta was detected, albeit in very small levels, by two-dimensional electrophoresis upon separation of PKAs in testes of 14-day-old rats by DEAE-cellulose chromatography but was absent in equivalent extracts from adult rat testes. These results demonstrate that the unexpectedly basic pI of RI beta allows for its clear separation by two-dimensional electrophoresis from the RII proteins and therefore allows for its unambiguous identification. Further studies, however, are required to resolve the basis for the apparent disparity in testis RI beta mRNA and protein.
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PMID:Characterization of recombinant RI beta and evaluation of the presence of RI beta protein in rat brain and testicular extracts. 803 21

Bovine seminal vesicles synthesize a family of closely related proteins, namely BSP-A1, BSP-A2, BSP-A3, and BSP-30-kDa (collectively called BSP proteins). Recently, we showed that these proteins bind specifically to choline phospholipids. Since this class of phospholipids is the major phospholipid fraction of the spermatozoan membrane, we investigated the binding of BSP proteins to spermatozoa. Polyclonal antibodies against purified BSP proteins raised in rabbits were used to detect these antigens in bovine epididymal and ejaculated spermatozoa as well as in bovine seminal plasma. Comparison of spermatozoa taken from the caudae epididymides with ejaculated spermatozoa through use of various techniques, namely, surface labeling followed by immunoprecipitation and immunoblotting, showed that epididymal spermatozoa are devoid of BSP proteins whereas ejaculated spermatozoa possess membrane-bound BSP proteins. Through use of the indirect immunofluorescence technique, the ejaculated spermatozoa of bull were characterized by an immunoreaction restricted to the midpiece, acrosome, and postacrosomal region, but no specific immunostaining could be found on the surface of epididymal spermatozoa. Surface-labeled BSP proteins on spermatozoa could not be displaced with buffers containing high salt concentration (1 M NaCl), but could be displaced specifically with phosphorylcholine (alone or in combination with urea). The data indicate that the BSP proteins that are secretory products of the seminal vesicles bind to the sperm surface upon ejaculation.
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PMID:Major proteins of bovine seminal vesicles bind to spermatozoa. 831 47

Chromatin of rat elongating spermatids, steps 12-13, is distinguished by the replacement of histones with transition proteins and the presence of nicks within its DNA which are formed by an endogenous nuclease, possibly DNA topoisomerase II (topo II). Using an affinity-purified anti-topo II antibody, protein bands of approximately 161 and approximately 137 kDa were detected on immunoblots of pachytene spermatocytes and elongating spermatids, respectively. In cryosections, topo II was localized to meiotic chromosomes of pachytene spermatocytes and to nuclei of elongating spermatids. Extracts of isolated testicular nuclei and sonication-resistant spermatid nuclei (steps 12-19) demonstrated topo II activity as determined by the decatenation of kinetoplast DNA. The potential relationship between nucleoprotein changes during spermatogenesis and the formation of nicks was also examined. Heterogeneous testicular and sonication-resistant spermatid nuclei were treated with 0.8 mM protamine, followed by nick translation in the absence of DNase I. In both cases, there was a dramatic decrease in DNA polymerase I-dependent label incorporation. To determine whether or not endogenous nicks were present in mature sperm, but were inaccessible due to protamine-DNA interactions, epididymal sperm were extracted with high salt-dithiothreitol, followed by nick translation in the absence or presence of DNase I. Extracted sperm nuclei did not nick translate in the absence of DNase I; however, incorporation increased with increasing concentrations of DNase I, indicating that endogenous nicks were repaired prior to the completion of spermatogenesis. These and previously published results suggest that topo II in elongating spermatids may be involved in the DNA alterations that take place during spermatogenesis, including changes in DNA topography, repair, and loop formation, and may serve as a component of the nuclear matrix. The temporal appearance and disappearance of endogenous nicks may reflect the changes that elongating spermatid DNA undergoes as a consequence of alterations in nucleoprotein composition to establish the condensed state of the mature spermatozoon.
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PMID:Nicking of rat spermatid and spermatozoa DNA: possible involvement of DNA topoisomerase II. 839 68

The relation was studied between the morphology of tests and epididymides on the one hand, and the lead content in these organs on the other. The testes of rats, which for the time of 5 spermatogeneses (9 months) were drinking 1% lead acetate(II), displayed all generations and layers of spermatogenic cells at respective stages of the seminiferous epithelium cycle. The lead content in testes of the animals did not differ significantly from the value of this element in gonads of control rats. The epididymal cells also failed to show morphological changes however, in all epididymal zones there were fewer spermatozoa than in the corresponding zones in control rats. Many spermatozoa revealed abnormal reactions of oxidoreductive enzymes in the midpiece. There were also local defects in tetrazolium salt reduction and segmental or total lack of formazan deposits in the mitochondrial sheath. The lead content in epididymes of these animals was significantly higher than in epididymides of control rats. The obtained results of the studies highlight the possibility for lead to accumulate in epididymis, and for lead compounds to damage spermatozoa in this organ.
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PMID:Effect of long-term exposure to lead on testis and epididymis in rats. 840 68

These studies were designed to assess the effects of pioglitazone, a new oral antidiabetic agent that acts by improving insulin sensitivity, on blood pressure, plasma and tissue lipids, and insulin resistance in the Dahl salt-sensitive (Dahl-S) rat. Reaven et al had reported that male Dahl-S rats are moderately hyperinsulinemic and insulin-resistant. This was of particular interest since these rats are not obese but are hypertriglyceridemic, and on a high-salt diet they become hypertensive. In the current study, male Sprague-Dawley control and Dahl-S rats were compared when fed standard chow of high-fat, high-sucrose (HFHS) diets with or without pioglitazone (20 mg/kg body weight/d) for 3 weeks. On the standard chow diet, Dahl-S rats were hypertriglyceridemic and had high tissue levels of malonyl coenzyme A ([CoA] Dahl-S 5.0 v control 3.3 nmol/g in muscle, and Dahl-S 15.6 v control 10.7 nmol/g in liver); however, they were not hyperinsulinemic. Pioglitazone therapy decreased both malonyl CoA and plasma triglycerides toward control values, but had no effect on plasma insulin levels. On the HFHS diet, both groups became glucose-intolerant and hyperinsulinemic; however, the hyperinsulinemia was greater and more sustained in Dahl-S rats. In addition, the HFHS diet appeared to increase the mass of retroperitoneal fat in the Dahl-S but not in the control group. Treatment with pioglitazone decreased retroperitoneal fat, but as reported previously, it increased the mass of the epididymal fat pad. The results suggest that the hypertriglyceridemia of the Dahl-S rat is associated with an increase in the concentration of malonyl CoA in both liver and muscle. They also show that pioglitazone reverses both of these abnormalities independently of its effect on plasma insulin. Whether these high levels of malonyl CoA predispose the Dahl-S rat to hyperinsulinemia and possibly obesity when placed on a HFHS diet remains to be determined.
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PMID:Malonyl coenzyme A and adiposity in the Dahl salt-sensitive rat: effects of pioglitazone. 860 42


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