A novel p64-related Cl−channel: subcellular distribution and nephron segment-specific expression

1999 ◽  
Vol 276 (3) ◽  
pp. F398-F408 ◽  
Author(s):  
John C. Edwards
Keyword(s):  
Chloride Channels ◽  
Sequence Similarity ◽  
Fluid Phase ◽  
Human Kidney ◽  
Specific Expression ◽  
Proximal Tubule Cells ◽  
Sds Page ◽  
Nephron Segment ◽  
Related Proteins

Several closely related proteins that have been implicated as chloride channels of intracellular membranes have recently been described. We report here the molecular cloning and characterization of a new member of this family from human cells. On the basis of sequence similarity, we conclude that this new protein represents the human version of a previously described protein from rat brain named p64H1. The human version of p64H1 (huH1) is a 28.7-kDa protein that shows an apparent molecular mass of 31 kDa by SDS-PAGE. A single 4.5-kb message is detected on Northern blots and is present in all tissues probed. The protein is expressed in an intracellular vesicular pattern in Panc-1 cells that is distinct from the endoplasmic reticulum, fluid-phase endocytic, and transferrin-recycling compartments, but which does colocalize with caveolin. In human kidney, huH1 is highly expressed in a diffuse pattern in the apical domain of proximal tubule cells. huH1 is expressed less abundantly in a vesicular pattern in glomeruli and distal nephron.

Toward a catalog for the transcripts and proteins (sialome) from the salivary gland of the malaria vectorAnopheles gambiae

2002 ◽  
Vol 205 (16) ◽  
pp. 2429-2451 ◽  
Author(s):  
Ivo M. B. Francischetti ◽  
Jesus G. Valenzuela ◽  
Van My Pham ◽  
Mark K. Garfield ◽  
José M. C. Ribeiro
Keyword(s):  
Salivary Gland ◽  
Sequence Similarity ◽  
Electronic Version ◽  
Hypothetical Proteins ◽  
Conserved Domain ◽  
Sds Page ◽  
Invaluable Tool ◽  
Related Proteins ◽  
Antigen 5

SUMMARYHundreds of Anopheles gambiae salivary gland cDNA library clones have been sequenced. A cluster analysis based on sequence similarity at e-60 grouped the 691 sequences into 251 different clusters that code for proteins with putative secretory, housekeeping, or unknown functions. Among the housekeeping cDNAs, we found sequences predicted to code for novel thioredoxin, tetraspanin, hemopexin, heat shock protein, and TRIO and MBF proteins. Among secreted cDNAs, we found 21 novel A. gambiaesalivary sequences including those predicted to encode amylase, calreticulin,selenoprotein, mucin-like protein and 30-kDa allergen, in addition to antigen 5- and D7-related proteins, three novel salivary gland (SG)-like proteins and eight unique putative secreted proteins (Hypothetical Proteins, HP). The electronic version of this paper contains hyperlinks to FASTA-formatted files for each cluster with the best match to the nonredundant (NR) and conserved domain databases (CDD) in addition to CLUSTAL alignments of each cluster. The N terminus of 12 proteins (SG-1, SG-1-like 2, SG-6, HP 8, HP 9-like, 5′nucleotidase, 30-kDa protein, antigen 5- and four D7-related proteins) has been identified by Edman degradation of PVDF-transferred, SDS/PAGE-separated salivary gland proteins. Therefore, we contribute to the generation of a catalog of A. gambiae salivary transcripts and proteins. These data are freely available and will eventually become an invaluable tool to study the role of salivary molecules in parasite-host/vector interactions.

Cloning and characterization of serpin-like genes from the striped rice stem borer, Chilo suppressalis

Genome ◽  
2013 ◽  
Vol 56 (6) ◽  
pp. 359-366 ◽  
Author(s):  
Zhao-Yu Ge ◽  
Pin-Jun Wan ◽  
Xiong-Feng Cheng ◽  
Yang Zhang ◽  
Guo-Qing Li ◽  
...  
Keyword(s):  
Proteinase Inhibitors ◽  
Sequence Similarity ◽  
Serine Proteinase ◽  
Stem Borer ◽  
Chilo Suppressalis ◽  
Specific Expression ◽  
Rice Stem Borer ◽  
Gut Physiology ◽  
The Impact

Serpins, also called serine proteinase inhibitors, are widely distributed in eukaryotes. In insects, serpins play important roles in regulating immune responses, gut physiology, and other processes. Here, we report the cloning and characterization of 12 serpin-like cDNAs from the striped rice stem borer (Chilo suppressalis), a major rice pest. The putative proteins share significant sequence similarity with known insect serpins, especially those from lepidopterons. Analysis of functional domains revealed that nine of the cloned serpins are putative trypsin- or chymotrypsin-like inhibitors; two are mixed-type serpins that may act as inhibitors for trypsins, elastases, or thrombin; and the remaining one is truncate. The potential functions of these serpins in interacting with host plants were also investigated by analyzing tissue-specific expression and the impact of different host plant genotypes on gene expression. Our results provide a foundation for future studies on the role of serpins in gut physiology in the striped rice stem borer, and also useful information for comparative analyses of serpins from different insect species.

scholarly journals Purification and characterization of cytosolic and microsomal cyclophilins from maize (Zea mays)

1996 ◽  
Vol 315 (3) ◽  
pp. 965-970 ◽  
Author(s):  
Philip S. SHELDON ◽  
Michael A. VENIS
Keyword(s):  
Cyclosporin A ◽  
Sequence Similarity ◽  
Terminal Sequence ◽  
Purification And Characterization ◽  
Cyclophilin B ◽  
Sds Page ◽  
Wide Range ◽  
Molecular Masses ◽  
High Level

Methods for the purification and separation of peptidyl prolyl cis–trans isomerase (PPI) from cytosolic and microsomal fractions of etiolated maize are described. On SDS/PAGE, the purified preparations appear as single polypeptides with molecular masses of 17.5 kDa and 17.7 kDa respectively. Instead of using immobilized cyclosporin A derivatives as affinity adsorbents, our methods employ conventional techniques enabling purification of the proteins on a much larger scale than previously described. An antiserum raised against the cytosolic PPI recognizes polypeptides of similar molecular mass from a wide range of plant species on an immunoblot. There is virtually no recognition of the microsomal PPI. The cytosolic and microsomal PPIs are inhibited by cyclosporin A (Ki = 6 nM in both cases), indicating that they are cyclophilins. The cytosolic enzyme is inactivated by 5 mM N-ethylmaleimide and 2 mM phenylglyoxal. N-terminal sequencing of the microsomal PPI indicates a high level of sequence similarity with the N-terminal sequence of mature animal s-cyclophilin (cyclophilin B).

Purification and characterization of activated human erythrocyte prolidase

1989 ◽  
Vol 67 (1) ◽  
pp. 34-41 ◽  
Author(s):  
Andrea M. Richter ◽  
Gerald L. Lancaster ◽  
Francis Y. M. Choy ◽  
Peter Hechtman
Keyword(s):  
Molecular Weight ◽  
Liquid Chromatography ◽  
Human Kidney ◽  
Human Erythrocytes ◽  
Rabbit Antibody ◽  
Purification And Characterization ◽  
Sds Page ◽  
Erythrocyte Enzyme ◽  
Chromatography Columns

Prolidase (E.C. 3.4.13.9) has been purified 7500-fold to homogeneity from human erythrocytes in a Mn2+-activated form using conventional and fast protein liquid chromatography columns. The procedure includes a 1-h incubation of the crude hemolysate at 50 °C with 1 mM MnCl2. Following this novel step, prolidase retains full activity, obviating the requirement for preincubation of each enzyme fraction with Mn2+ prior to assay. Preincubation with MnCl2 does not change the isoelectric point of the enzyme. The molecular weight of the purified enzyme was 58 000 when measured by SDS–PAGE. Western blotting, using rabbit antibody raised to human kidney prolidase, with partially purified erythrocyte enzyme revealed a cross-reacting band at Mr 58 000.Key words: prolidase purification, human erythrocytes.

scholarly journals Purification and partial characterization of rat factor D

1991 ◽  
Vol 279 (3) ◽  
pp. 775-779 ◽  
Author(s):  
B C Baker ◽  
C J Campbell ◽  
C J Grinham ◽  
G Turcatti
Keyword(s):  
Wheat Germ ◽  
Human Factor ◽  
Sequence Similarity ◽  
Terminal Sequence ◽  
Sds Page ◽  
Fast Flow ◽  
Sepharose 4B ◽  
Wheat Germ Lectin ◽  
Human And Mouse

Rat factor D has been purified to homogeneity (10,559-fold) from serum by chromatography on CM-Sepharose Fast Flow, phenyl-Sepharose CL-4B and Mono S and has been shown to resemble its human and mouse counterparts both in substrate specificity and in its susceptibility to inhibition by the organophosphorous inhibitor di-isopropylfluorophosphate. The rat enzyme, however, is heavily glycosylated and binds to wheat-germ lectin-Sepharose 6MB and 5-hydroxytryptamine-agarose, but not to concanavalin A-Sepharose 4B. All of the carbohydrate chains are N-linked. Enzymic removal of this carbohydrate decreased the Mr by approx. 15,000. The deglycosylated rat enzyme had the same mobility as native human factor D on SDS/PAGE, corresponding to an Mr of 24,500. N-Terminal sequence analysis of the first 30 amino acids of rat factor D highlighted the sequence similarity with human factor D (greater than 76%) and, in particular, with mouse adipsin (greater than 93%).

scholarly journals Telescope: Characterization of the retrotranscriptome by accurate estimation of transposable element expression

2018 ◽  
Author(s):  
Matthew L. Bendall ◽  
Miguel de Mulder ◽  
Luis Pedro Iñiguez ◽  
Aarón Lecanda-Sánchez ◽  
Marcos Pérez-Losada ◽  
...  
Keyword(s):  
Transposable Element ◽  
Sequence Similarity ◽  
Cell Types ◽  
Accurate Estimation ◽  
Rna Seq ◽  
Cell Type ◽  
Specific Expression ◽  
Probable Source ◽  
Herv Expression

AbstractCharacterization of Human Endogenous Retrovirus (HERV) expression within the transcriptomic landscape using RNA-seq is complicated by uncertainty in fragment assignment because of sequence similarity. We present Telescope, a computational software tool that provides accurate estimation of transposable element expression (retrotranscriptome) resolved to specific genomic locations. Telescope directly addresses uncertainty in fragment assignment by reassigning ambiguously mapped fragments to the most probable source transcript as determined within a Bayesian statistical model. We demonstrate the utility of our approach through single locus analysis of HERV expression in 13 ENCODE cell types. When examined at this resolution, we find that the magnitude and breadth of the retrotranscriptome can be vastly different among cell types. Furthermore, our approach is robust to differences in sequencing technology, and demonstrates that the retrotranscriptome has potential to be used for cell type identification. Telescope performs highly accurate quantification of the retrotranscriptomic landscape in RNA-seq experiments, revealing a differential complexity in the transposable element biology of complex systems not previously observed. Telescope is available at github.com/mlbendall/telescope.Author SummaryAlmost half of the human genome is composed of Transposable elements (TEs), but their contribution to the transcriptome, their cell-type specific expression patterns, and their role in disease remains poorly understood. Recent studies have found many elements to be actively expressed and involved in key cellular processes. For example, human endogenous retroviruses (HERVs) are reported to be involved in human embryonic stem cell differentiation. Discovering which exact HERVs are differentially expressed in RNA-seq data would be a major advance in understanding such processes. However, because HERVs have a high level of sequence similarity it is hard to identify which exact HERV is differentially expressed. To solve this problem, we developed a computer program which addressed uncertainty in fragment assignment by reassigning ambiguously mapped fragments to the most probable source transcript as determined within a Bayesian statistical model. We call this program, “Telescope”. We then used Telescope to identify HERV expression in 13 well-studied cell types from the ENCODE consortium and found that different cell types could be characterized by enrichment for different HERV families, and for locus specific expression. We also showed that Telescope performed better than other methods currently used to determine TE expression. The use of this computational tool to examine new and existing RNA-seq data sets may lead to new understanding of the roles of TEs in health and disease.

Characterization of embryogenesis-related proteins in alfalfa (Medicago sativa)

1996 ◽  
Vol 96 (4) ◽  
pp. 585-592 ◽  
Author(s):  
Randal W. Giroux ◽  
K. Peter Pauls
Keyword(s):  
Medicago Sativa ◽  
Related Proteins

Characterization of Venoms of Deinagkistrodon acutus and Bungarus multicinctus Using Proteomics and Peptidomics

2020 ◽  
Vol 17 (3) ◽  
pp. 241-254
Author(s):  
Yaqiong Zhang ◽  
Zhiping Jia ◽  
Yunyang Liu ◽  
Xinwen Zhou ◽  
Yi Kong
Keyword(s):  
Snake Venom ◽  
Protein Families ◽  
Traditional Medicines ◽  
Phospholipases A2 ◽  
Inhibitory Peptides ◽  
Sds Page ◽  
Deinagkistrodon Acutus ◽  
Venom Proteins ◽  
Proteins And Peptides

Background: Deinagkistrodon acutus (D. acutus) and Bungarus multicinctus (B. multicinctus) as traditional medicines have been used for hundreds of years in China. The venoms of these two species have strong toxicity on the victims. Objective: The objective of this study is to reveal the profile of venom proteins and peptides of D. acutus and B. multicinctus. Method: Ultrafiltration, SDS-PAGE coupled with in-gel tryptic digestion and Liquid Chromatography- Electrospray Ionization-Tandem Mass Spectrometry (LC-ESI-MS/MS) were used to characterize proteins and peptides of venoms of D. acutus and B. multicinctus. Results: In the D. acutus venom, 67 proteins (16 protein families) were identified, and snake venom metalloproteinases (SVMPs, 38.0%) and snake venom C-type lectins (snaclecs, 36.7%) were dominated proteins. In the B. multicinctus venom, 47 proteins (15 protein families) were identified, and three-finger toxins (3FTxs, 36.3%) and Kunitz-type Serine Protease Inhibitors (KSPIs, 32.8%) were major components. In addition, both venoms contained small amounts of other proteins, such as Snake Venom Serine Proteinases (SVSPs), Phospholipases A2 (PLA2s), Cysteine-Rich Secreted Proteins (CRISPs), 5'nucleotidases (5'NUCs), Phospholipases B (PLBs), Phosphodiesterases (PDEs), Phospholipase A2 Inhibitors (PLIs), Dipeptidyl Peptidases IV (DPP IVs), L-amino Acid Oxidases (LAAOs) and Angiotensin-Converting Enzymes (ACEs). Each venom also had its unique proteins, Nerve Growth Factors (NGFs) and Hyaluronidases (HYs) in D. acutus, and Cobra Venom Factors (CVFs) in B. multicinctus. In the peptidomics, 1543 and 250 peptides were identified in the venoms of D. acutus and B. multicinctus, respectively. Some peptides showed high similarity with neuropeptides, ACE inhibitory peptides, Bradykinin- Potentiating Peptides (BPPs), LAAOs and movement related peptides. Conclusion: Characterization of venom proteins and peptides of D. acutus and B. multicinctus will be helpful for the treatment of envenomation and drug discovery.

Characterization of Glycoprotein Fraction from Carp Pituitaries Isolated Using Concanavalin A as the Affinity Ligand

1998 ◽  
Vol 63 (3) ◽  
pp. 434-440 ◽  
Author(s):  
Irena Hulová ◽  
Jana Barthová ◽  
Helena Ryšlavá ◽  
Václav Kašička
Keyword(s):  
Concanavalin A ◽  
Reversed Phase ◽  
Amino Acid Sequences ◽  
Gel Chromatography ◽  
Affinity Ligand ◽  
Sds Page ◽  
Terminal Amino ◽  
Α And Β Subunits

Glycoproteins that have affinity to Concanavalin A were isolated from the acetone-dried pituitaries of common carp (Cyprinus carpio L.). Two fractions of glycoproteins were separated using gel chromatography on Superdex 75HR. The fraction with lower molecular weight (30 000) corresponding to the carp gonadotropin cGtH II was composed of two subunits as determined using SDS-PAGE. This protein fraction was further divided into four components using reversed-phase HPLC. Two fractions were pure α and β subunits of cGtH II as follows from immunodetection and from determination of N-terminal amino acid sequences. The other two were a mixture of α and β subunits as was also revealed by N-terminal analysis. Capillary electrophoresis was also used for characterization of isolated glycoproteins.

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