scholarly journals A novel testis-specific protein, PRAMEY, is involved in spermatogenesis in cattle

Reproduction ◽  
2017 ◽  
Vol 153 (6) ◽  
pp. 847-863 ◽  
Author(s):  
Wan-Sheng Liu ◽  
Yaqi Zhao ◽  
Chen Lu ◽  
Gang Ning ◽  
Yun Ma ◽  
...  
Keyword(s):  
Specific Protein ◽  
Immunofluorescent Staining ◽  
Immunogold Electron Microscopy ◽  
Spermatogenic Cells ◽  
Mature Protein ◽  
Chinese Translation ◽  
Link Type ◽  
The Matrix ◽  
Epididymal Spermatozoa ◽  
Cancer Testis

Preferentially expressed antigen in melanoma (PRAME) is a cancer/testis antigen that is predominantly expressed in normal testicular tissues and a variety of tumors. The function of the PRAME family in spermatogenesis remains unknown. This study was designed to characterize the Y-linked PRAME (PRAMEY) protein during spermatogenesis in cattle. We found that PRAMEY is a novel male germ cell-specific, and a germinal granule-associated protein that is expressed in spermatogenic cells during spermatogenesis. The intact PRAMEY protein (58 kDa) was detected in different ages of testes but not in epididymal spermatozoa. A PRAMEY isoform (30 kDa) was highly expressed only in testes after puberty and in epididymal spermatozoa. This isoform interacts with PP1γ2 and is likely the mature protein present in the testes and sperm. Immunofluorescent staining demonstrated that PRAMEY was located predominantly in the acrosome granule of spermatids, and in acrosome and flagellum of spermatozoa. Immunogold electron microscopy further localized the PRAMEY protein complex to the nucleus and several cytoplasmic organelles, including the rough endoplasmic reticulum, some small vesicles, the intermitochondrial cement, the chromatoid body and the centrioles, in spermatogonia, spermatocytes, spermatids and/or spermatozoa. PRAMEY was highly enriched in and structurally associated with the matrix of the acrosomal granule (AG) in round spermatids, and migrated with the expansion of the AG during acrosomal biogenesis. While the function of PRAMEY remains unclear during spermatogenesis, our results suggest that PRAMEY may play an essential role in acrosome biogenesis and spermatogenesis.Free Chinese abstract: A Chinese translation of this abstract is freely available athttp://www.reproduction-online.org/content/153/6/847/suppl/DC2Spanish abstract: A Spanish translation of this abstract is freely available athttp://www.reproduction-online.org/content/153/6/847/suppl/DC3

scholarly journals von Willebrand factor released from Weibel-Palade bodies binds more avidly to extracellular matrix than that secreted constitutively

Blood ◽  
1987 ◽  
Vol 69 (5) ◽  
pp. 1531-1534 ◽  
Author(s):  
LA Sporn ◽  
VJ Marder ◽  
DD Wagner
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Von Willebrand Factor ◽  
Immunofluorescent Staining ◽  
Cultured Endothelial Cells ◽  
Human Foreskin Fibroblasts ◽  
Platelet Binding ◽  
The Matrix ◽  
Von Willebrand ◽  
Willebrand Factor

Abstract Large multimers of von Willebrand factor (vWf) are released from the Weibel-Palade bodies of cultured endothelial cells following treatment with a secretagogue (Sporn et al, Cell 46:185, 1986). These multimers were shown by immunofluorescent staining to bind more extensively to the extracellular matrix of human foreskin fibroblasts than constitutively secreted vWf, which is composed predominantly of dimeric molecules. Increased binding of A23187-released vWf was not due to another component present in the releasate, since releasate from which vWf was adsorbed, when added together with constitutively secreted vWf, did not promote binding. When iodinated plasma vWf was overlaid onto the fibroblasts, the large forms bound preferentially to the matrix. These results indicated that the enhanced binding of the vWf released from the Weibel-Palade bodies was likely due to its large multimeric size. It appears that multivalency is an important component of vWf interaction with the extracellular matrix, just as has been shown for vWf interaction with platelets. The pool of vWf contained within the Weibel-Palade bodies, therefore, is not only especially suited for platelet binding, but also for interaction with the extracellular matrix.

Spinalin, a new glycine- and histidine-rich protein in spines of Hydra nematocysts

1998 ◽  
Vol 111 (11) ◽  
pp. 1545-1554 ◽  
Author(s):  
A.W. Koch ◽  
T.W. Holstein ◽  
C. Mala ◽  
E. Kurz ◽  
J. Engel ◽  
...  
Keyword(s):  
External Surface ◽  
Insoluble Fraction ◽  
Cdna Clones ◽  
Mature Protein ◽  
Capsule Wall ◽  
The Matrix ◽  
Acidic Region ◽  
Basic Region ◽  
Full Length Clone

Here we present the cloning, expression and immunocytochemical localization of a novel 24 kDa protein, designated spinalin, which is present in the spines and operculum of Hydra nematocysts. Spinalin cDNA clones were identified by in situ hybridization to differentiating nematocytes. Sequencing of a full-length clone revealed the presence of an N-terminal signal peptide, suggesting that the mature protein is sorted via the endoplasmic reticulum to the post-Golgi vacuole in which the nematocyst is formed. The N-terminal region of spinalin (154 residues) is very rich in glycines (48 residues) and histidines (33 residues). A central region of 35 residues contains 19 glycines, occurring mainly as pairs. For both regions a polyglycine-like structure is likely and this may be stabilized by hydrogen bond-mediated chain association. Similar sequences found in loricrins, cytokeratins and avian keratins are postulated to participate in formation of supramolecular structures. Spinalin is terminated by a basic region (6 lysines out of 15 residues) and an acidic region (9 glutamates and 9 aspartates out of 32 residues). Western blot analysis with a polyclonal antibody generated against a recombinant 19 kDa fragment of spinalin showed that spinalin is localized in nematocysts. Following dissociation of the nematocyst's capsule wall with DTT, spinalin was found in the insoluble fraction containing spines and the operculum. Immunocytochemical analysis of developing nematocysts revealed that spinalin first appears in the matrix but then is transferred through the capsule wall at the end of morphogenesis to form spines on the external surface of the inverted tubule and the operculum.

Removal of a hydrophobic domain within the mature portion of a mitochondrial inner membrane protein causes its mislocalization to the matrix

1990 ◽  
Vol 10 (5) ◽  
pp. 1873-1881
Author(s):  
S M Glaser ◽  
B R Miller ◽  
M G Cumsky
Keyword(s):  
Amino Acids ◽  
Mitochondrial Matrix ◽  
Mature Protein ◽  
Wild Type ◽  
Inner Membrane ◽  
Hydrophobic Domain ◽  
Mitochondrial Inner Membrane ◽  
The Matrix ◽  
Inner Membrane Protein ◽  
Cognate Protein

We have examined the import and intramitochondrial localization of the precursor to yeast cytochrome c oxidase subunit Va, a protein of the mitochondrial inner membrane. The results of studies on the import of subunit Va derivatives carrying altered presequences suggest that the uptake of this protein is highly efficient. We found that a presequence of only 5 amino acids (Met-Leu-Ser-Leu-Arg) could direct the import and localization of subunit Va with wild-type efficiency, as judged by several different assays. We also found that subunit Va could be effectively targeted to the mitochondrial inner membrane with a heterologous presequence that failed to direct import of its cognate protein. The results presented here confirmed those of an earlier study and showed clearly that the information required to "sort" subunit Va to the inner membrane resides in the mature protein sequence, not within the presequence per se. We present additional evidence that the aforementioned sorting information is contained, at least in part, in a hydrophobic stretch of 22 amino acids residing within the C-terminal third of the protein. Removal of this domain caused subunit Va to be mislocalized to the mitochondrial matrix.

scholarly journals Heterogeneity in the pattern of distribution of the specific hormonal product and secretogranins within the secretory granules of rat prolactin cells.

1994 ◽  
Vol 42 (8) ◽  
pp. 1097-1107 ◽  
Author(s):  
H Ozawa ◽  
R Picart ◽  
A Barret ◽  
C Tougard
Keyword(s):  
Subcellular Distribution ◽  
Secretory Granules ◽  
Culture Conditions ◽  
Male Rat ◽  
Immunogold Electron Microscopy ◽  
The Matrix ◽  
Granule Membrane ◽  
Adult Male Rat ◽  
Secretory Products ◽  
Rat Pituitary Tumor Cells

We investigated the subcellular distribution of secretogranins I, II (Sg I, Sg II), and prolactin (PRL) by double immunogold electron microscopy in GH3B6 rat pituitary tumor cells grown in different culture conditions and in normal PRL cells in adult male rat anterior pituitary. Co-localization of Sg I or Sg II with PRL was observed in most secretory granules in GH3B6 cells and normal PRL cells, except for some secretory granules containing only Sgs in GH3B6 cells and containing only PRL in normal PRL cells. In GH3B6 cells treated with thyrotropin-releasing hormone (TRH) for 2 hr, the newly formed small secretory granules within the Golgi zone contained preferentially immunoreactive PRL. Interestingly, when co-localized with PRL, Sgs (particularly Sg I) were observed at the periphery of the matrix of secretory granules in GH3B6 cells as well as in normal PRL cells, suggesting their possible interaction with the secretory granule membrane. The present study indicates a heterogeneous subcellular distribution of PRL, Sg I, and Sg II in individual secretory granules of GH3B6 cells and of normal PRL cells, pointing out the formation of different types of aggregates during the condensation of secretory products in these two PRL cell models.

scholarly journals Meixner polynomials of the second kind and quantum algebras representing su(1, 1)

2009 ◽  
Vol 466 (2117) ◽  
pp. 1409-1428 ◽  
Author(s):  
Gábor Hetyei
Keyword(s):  
Orthogonal Polynomials ◽  
Laguerre Polynomials ◽  
Combinatorial Theory ◽  
Quantum Algebras ◽  
Combinatorial Approach ◽  
Meixner Polynomials ◽  
Link Type ◽  
Polynomial Coefficients ◽  
The Matrix ◽  
Math Phys

We show how Viennot’s combinatorial theory of orthogonal polynomials may be used to generalize some recent results of Sukumar and Hodges (Hodges & Sukumar 2007 Proc. R. Soc. A 463 , 2401–2414 ( doi:10.1098/rspa.2007.0001 ); Sukumar & Hodges 2007 Proc. R. Soc. A 463 , 2415–2427 ( doi:10.1098/rspa.2007.0003 )) on the matrix entries in powers of certain operators in a representation of su(1, 1). Our results link these calculations to finding the moments and inverse polynomial coefficients of certain Laguerre polynomials and Meixner polynomials of the second kind. As an immediate consequence of results by Koelink, Groenevelt and Van Der Jeugt (Van Der Jeugt 1997 J. Math. Phys. 38 , 2728–2740 ( doi:10.1063/1.531984 ); Koelink & Van Der Jeugt 1998 SIAM J. Math. Anal. 29 , 794–822 ( doi:10.1137/S003614109630673X ); Groenevelt & Koelink 2002 J. Phys. A 35 , 65–85 ( doi:10.1088/0305-4470/35/1/306 )), for the related operators, substitutions into essentially the same Laguerre polynomials and Meixner polynomials of the second kind may be used to express their eigenvectors. Our combinatorial approach explains and generalizes this ‘coincidence’.

scholarly journals Spectrum of pathogenic variants and multiple founder effects in amelogenesis imperfecta associated with MMP20

2020 ◽  
Author(s):  
Georgios Nikolopoulos ◽  
Claire E. L. Smith ◽  
James A. Poulter ◽  
Gina Murillo ◽  
Sandra Silva ◽  
...  
Keyword(s):  
Protein Function ◽  
Mendelian Inheritance ◽  
Data Availability ◽  
Amelogenesis Imperfecta ◽  
Compound Heterozygous ◽  
Link Type ◽  
Pathogenic Variants ◽  
The Matrix ◽  
Variation Database ◽  
Probable Impact

AbstractAmelogenesis imperfecta (AI) describes a heterogeneous group of developmental enamel defects that typically have Mendelian inheritance. Exome sequencing of ten families with recessive hypomaturation AI revealed 4 novel and 1 known variants in the matrix metallopeptidase 20 (MMP20) gene that were predicted to be pathogenic. MMP20 encodes a protease that cleaves the developing extracellular enamel matrix and is necessary for normal enamel crystal growth during amelogenesis. New homozygous missense changes were shared between four families of Pakistani heritage (c.625G>C; p.(E209Q)) and two of Omani origin (c.710C>A; p.(S237Y)). In two families of UK origin and one from Costa Rica, affected individuals were homozygous for the previously reported c.954-2A>T; p.(I319Ffs*19) variant. For each of these variants, microsatellite haplotypes appeared to exclude a recent founder effect, but elements of haplotype were conserved, suggesting more distant founding ancestors. New compound heterozygous changes were identified in one family of European heritage; c.809_811+12delACGgtaagattattainsCCAG; p.(?) and c.1122A>C; p.(Q374H). All four new variants are within the zinc dependant peptidase domain. This report further elucidates the mutation spectrum of MMP20 and the probable impact on protein function, confirms a consistent hypomaturation phenotype and shows that mutations in MMP20 are a common cause of autosomal recessive AI in some communities.Data AvailabilityThe data that support the findings of this study are openly available in ClinVar at https://www.ncbi.nlm.nih.gov/clinvar/, accession numbers: SCV001338799 - SCV001338802 and in the AI Leiden Open Variation Database (LOVD) at http://dna2.leeds.ac.uk/LOVD/ with reference numbers: 0000000313 – 0000000317.

scholarly journals Monoclonal Antibodies to a Subfraction of Merino Wool High-tyrosine Proteins

1986 ◽  
Vol 39 (4) ◽  
pp. 341 ◽  
Author(s):  
DR Hewish ◽  
PW French
Keyword(s):  
Electron Microscopy ◽  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
Immunofluorescent Staining ◽  
Immunogold Electron Microscopy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Location ◽  
Merino Wool

Monoclonal antibodies were prepared which react with members of the high-tyrosine type proteins from Merino wool. Specificity was confirmed by the use of Western transfer immunoassays and by enzyme-linked immunosorbent assay on purified fractions. Immunofluorescent staining of sections of wool follicles using the antibodies showed that the proteins were present in the developing wool shaft but that staining was asymmetric, indicating specific location of the proteins in the orthocortex of the fibres. Immunogold-electron microscopy confirmed that one of the antibodies bound to the keratin microfibril bundles.

scholarly journals Analysis of epitope distribution of arabinogalactan protein-extensins in pea (Pisum Sativum) nodules of wild-type and mutants impaired in infection thread growth

2019 ◽  
Vol 17 (3) ◽  
pp. 5-12
Author(s):  
Anna V. Tsyganova ◽  
Nicholas J. Brewin ◽  
Viktor E. Tsyganov
Keyword(s):  
Mutant Line ◽  
Arabinogalactan Protein ◽  
Infection Thread ◽  
Immunogold Electron Microscopy ◽  
Bacterial Cells ◽  
Wild Type ◽  
Infection Threads ◽  
The Matrix ◽  
Pea Mutants ◽  
High Level

Background. During the colonization of root and nodule tissues of legumes by rhizobia, bacterial cells are immersed in a plant extracellular matrix which includes arabinogalactan protein-extensins (AGPE). Materials and methods. Immunogold electron microscopy with monoclonal antibodies MAC204 and MAC236 was used to analyse the distribution and abundance of epitopes of AGPE in wild-type and symbiotically defective pea mutants. Results. In the nodules of the wild-type line SGE, both AGPE epitopes were detected to the same extent in the matrix of infection threads and infection droplets. In the nodules of the mutant line SGEFix-1 (sym40), the level of labelling by MAC204 was significantly higher than with SGE in both infection threads and infection droplets, but the level of labelling by MAC236 was only increased in the infection droplets. In the mutant line SGEFix-2 (sym33-3), a relatively high level of both epitopes was observed among all analysed genotypes. The double mutant line RBT3 (sym33-3, sym40) showed an intermediate level of labelling for both epitopes in infection threads compared with the parental mutants. In SGEFix-1, an abnormal distribution of both epitopes was observed in the intercellular space matrix. The MAC204 epitope was found in the cell walls of SGEFix-1 and in the infection thread walls of SGEFix-2, whereas in RBT3 this epitope was detected in both types of walls. Conclusions. The sym33-3 and sym40 mutations have different effects on the accumulation of AGPE epitopes recognised by MAC204 and MAC236. This indicates that both the Sym33 and the Sym40 genes affect the composition of AGPE in the matrix of infection threads and infection droplets.

scholarly journals Immunohistochemical Study of Thrombolytic Mechanisms

Blood ◽  
1965 ◽  
Vol 25 (6) ◽  
pp. 1028-1036 ◽  
Author(s):  
N. BACK ◽  
R. HIRAMOTO ◽  
J. L. AMBRUS
Keyword(s):  
Immunohistochemical Study ◽  
Fibrinolytic Enzyme ◽  
Immunofluorescent Staining ◽  
Enzyme Preparations ◽  
The Matrix ◽  
Human Plasminogen ◽  
High Degree ◽  
Indirect Immunofluorescent ◽  
Fibrin Clots

Abstract The indirect immunofluorescent staining technic has been employed to study the localization onto fibrin clots of components of the fibrinolysin system and its activators. Immunodiffusion technics revealed the heterogeneity of the various enzyme preparations used. The activated fibrinolytic enzyme preparations were found to localize onto and diffuse into the matrix and core of the clots. High degree of localization was seen with streptokinase-, urokinase- and spontaneously activated human plasmin, as well as human plasminogen. Chloroform-activated bovine plasmin localized to a lesser extent. No differences were observed in the results whether the fibrin clots were of human, canine or bovine origin.

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