scholarly journals Identification of a filamentous form of kunitz protease inhibitor in Asteraceae

2021 ◽  
Author(s):  
Sara Bratsch ◽  
Neil Olszewski ◽  
Benham Lockhart
Keyword(s):  
Protease Inhibitor ◽  
Protein Sequencing ◽  
Major Protein ◽  
Kunitz Trypsin Inhibitor ◽  
Terminal Protein ◽  
Filamentous Form ◽  
First Report ◽  
Kunitz Protease Inhibitor ◽  
Sds Page ◽  
Filament Protein

AbstractFilamentous structures were observed in purified extracts from chrysanthemum, gerbera, sunflower and zinnia. When purified filament proteins were subjected to SDS-PAGE, the major protein associated with filaments from all three species has an apparent molecular mass of ≈25 kDa. Protein bands from chrysanthemum, gerbera, and zinnia were subjected to N-terminal protein sequencing while proteins from sunflower were sequenced by CID MS/MS. All of the sequences shared highest similarity to the kunitz trypsin inhibitor family. The sequencing results indicated that the proteins lacked the signal sequences. We tested the gerbera filament protein for glycosylation and found that it was a glycoprotein. Together these results indicate that the filaments are composed of mature KTI protein. This is the first report of a KTI assembling into filaments and the first report of a filament forming Asteraceae enzyme.

scholarly journals First Report of the Cymbidium Mosaic Potexvirus (CymMV) Infecting the Terrestrial Orchid Phaius tankervilliae in Costa Rica

Plant Disease ◽  
1998 ◽  
Vol 82 (10) ◽  
pp. 1171-1171 ◽  
Author(s):  
L. Moreira ◽  
W. Villalobos ◽  
H. T. Hsu ◽  
E. Rodríguez-Cerezo ◽  
C. Rivera
Keyword(s):  
Costa Rica ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Central Valley ◽  
Major Protein ◽  
Terrestrial Orchid ◽  
First Report ◽  
Foliar Symptoms ◽  
Viral Particles ◽  
Sds Page

In 1996, plants of the terrestrial orchid Phaius tankervilliae from a nursery in the Central Valley of Costa Rica were observed with mild to severe foliar symptoms of chlorotic streak. No differences were observed in growth, bulb production, flowers, or flowering time between symptomatic and asymptomatic plants, except the symptomatic plants had earlier senescence. Occasionally, the flowers displayed symptoms of chlorosis and white rings in the sepals. Extracts from symptomatic leaves were concentrated by differential centrifugation and analyzed after sucrose gradients. Negative staining of fractions from gradients from symptomatic plants showed the presence of filamentous viral particles 500 by 17 nm. Purified particles contained a single major protein of about 28 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and a single RNA of about 7 kb, which is greater than the 6.2 kb reported (GenBank). These data suggest the presence of a potexvirus in symptomatic plants (1,2). In enzyme-linked immunosorbent assays, symptomatic plants reacted strongly with antiserum specific for Cymbidium mosaic potexvirus (CymMV). This is the first report of CymMV in P. tankervilliae in Costa Rica. References: (1) J. A. Frowd and J. H. Tremaine. Phytopathology 67:43, 1977. (2) H. T. Hsu et al. Phytopathology 82:491, 1992.

scholarly journals Intranuclear filaments containing a nuclear pore complex protein.

1993 ◽  
Vol 123 (6) ◽  
pp. 1333-1344 ◽  
Author(s):  
V C Cordes ◽  
S Reidenbach ◽  
A Köhler ◽  
N Stuurman ◽  
R van Driel ◽  
...  
Keyword(s):  
Nucleocytoplasmic Transport ◽  
Soluble Form ◽  
Molecular Entity ◽  
Nuclear Pore ◽  
Nuclear Pore Complexes ◽  
Filamentous Form ◽  
Sds Page ◽  
Complex Protein ◽  
Pore Complexes ◽  
Filament Protein

Nuclear pore complexes (NPCs) are anchoring sites of intranuclear filaments of 3-6 nm diameter that are coaxially arranged on the perimeter of a cylinder and project into the nuclear interior for lengths varying in different kinds of cells. Using a specific monoclonal antibody we have found that a polypeptide of approximately 190 kD on SDS-PAGE, which appears to be identical to the recently described NPC protein "nup 153," is a general constituent of these intranuclear NPC-attached filaments in different types of cells from diverse species, including amphibian oocytes where these filaments are abundant and can be relatively long. We have further observed that during mitosis this filament protein transiently disassembles, resulting in a distinct soluble molecular entity of approximately 12.5 S, and then disperses over most of the cytoplasm. Similarly, the amphibian oocyte protein appears in a soluble form of approximately 16 S during meiotic metaphase and can be immunoprecipitated from egg cytoplasmic supernatants. We conclude that this NPC protein can assemble into a filamentous form at considerable distance from the nuclear envelope and discuss possible functions of these NPC-attached filaments, from a role as guidance structure involved in nucleocytoplasmic transport to a form of excess storage of NPC proteins in oocytes.

P4-153: The disruption of A-Beta peptide by transthyretin: A mechanism sensitive to Kunitz Protease Inhibitor

2009 ◽  
Vol 5 (4S_Part_16) ◽  
pp. P476-P476
Author(s):  
Rita Costa ◽  
Frederico Ferreira-da Silva ◽  
Maria Joao Saraiva ◽  
Isabel Cardoso
Keyword(s):  
Protease Inhibitor ◽  
Kunitz Protease Inhibitor ◽  
Beta Peptide

A metalloproteinase extracellularly released byCrithidia deanei

2003 ◽  
Vol 49 (10) ◽  
pp. 625-632 ◽  
Author(s):  
Claudia Masini d'Avila-Levy ◽  
Rodrigo F Souza ◽  
Rosana C Gomes ◽  
Alane B Vermelho ◽  
Marta H Branquinha
Keyword(s):  
Molecular Mass ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Residual Activity ◽  
Major Protein ◽  
Motile Cells ◽  
Sds Page ◽  
Triton X

Actively motile cells from a cured strain of Crithidia deanei released proteins in phosphate buffer (pH 7.4). The molecular mass of the released polypeptides, which included some proteinases, ranged from 19 to 116 kDa. One of the major protein bands was purified to homogeneity by a combination of anion-exchange and gel filtration chromatographs. The apparent molecular mass of this protein was estimated to be 62 kDa by sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS–PAGE). The incorporation of gelatin into SDS–PAGE showed that the purified protein presented proteolytic activity in a position corresponding to a molecular mass of 60 kDa. The enzyme was optimally active at 37 °C and pH 6.0 and showed 25% of residual activity at 28 °C for 30 min. The proteinase was inhibited by 1,10-phenanthroline and EDTA, showing that it belonged to the metalloproteinase class. A polyclonal antibody to the leishmanial gp63 reacted strongly with the released C. deanei protease. After Triton X-114 extraction, an enzyme similar to the purified metalloproteinase was detected in aqueous and detergent-rich phases. The detection of an extracellular metalloproteinase produced by C. deanei and some other Crithidia species suggests a potential role of this released enzyme in substrate degradation that may be relevant to the survival of trypanosomatids in the host.Key words: endosymbiont, trypanosomatid, extracellular, proteinase.

Proteomics Variation in Nigella Sativa L. (Rananculaceae) Germplasm

2021 ◽  
Vol 50 (2) ◽  
pp. 289-294
Author(s):  
Muhammad Sajjad Iqbal ◽  
Abdul Ghafoor
Keyword(s):  
Euclidean Distance ◽  
Nigella Sativa ◽  
Crop Improvement ◽  
Band Formation ◽  
First Report ◽  
Sds Page ◽  
Genetic Pattern ◽  
Specific Variation ◽  
Improvement Programs ◽  
Single Seeds

Study revealed a first report of proteomics variation in Nigella sativa L. based on analyzing 32 accessions through SDS-PAGE. Three prominent regions along eight subunits were identified. Intra specific variation was observed low whereas the sharpness of bands was high between first and second regions. It was noted that in second region there was no clear evidence of band formation in N. sativa. Prominent and sharp protein peptide bands were recorded in four accessions, namely PK-020561, PK-020609, PK-020620 and PK-020646. Further investigation of single seeds showed almost similar genetic pattern within the single accession. Five clusters were formed on the basis of Euclidean distance. Cluster-I & II contain 1, 1 accession each, likewise Cluster-III and C-IV contain 2, 2 accessions whereas Cluster-V was found diversified as consisted of 26 accessions. Two accessions PK-020878 and PK-020877 were recommended for polymorphism and crop improvement programs. Bangladesh J. Bot. 50(2): 289-294, 2021 (June)

Cytoplasmic surface and intramembrane components of rat heart gap junctional proteins

1984 ◽  
Vol 246 (6) ◽  
pp. H865-H875 ◽  
Author(s):  
C. K. Manjunath ◽  
G. E. Goings ◽  
E. Page
Keyword(s):  
Electron Microscopy ◽  
Gap Junctions ◽  
Electron Micrographs ◽  
Rat Heart ◽  
Major Protein ◽  
Thin Sections ◽  
Cytoplasmic Surface ◽  
Sds Page ◽  
Junction Protein ◽  
Major Protein Band

Gap junctions were purified from rat hearts in the presence of absence of proteolysis inhibitors and examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and electron microscopy of thin sections. In absence of proteolysis inhibitors or in presence of ethylenediaminetetraacetic acid or leupeptin, gap junctions contained a single major protein band at relative molecular weight (Mr) 29,500 and minor bands at Mr 44,000–47,000, 17,750, and 16,500 and showed smooth cytoplasmic surfaces in electron micrographs. SDS-PAGE of junctions prepared with phenylmethylsulfonylfluoride (PMSF) showed markedly decreased intensity of the Mr 29,500 band and increased intensity of bands at Mr 44,000, 45,500, and 47,000; electron microscopy of these gap junctions showed presence of a fuzzy layer on their cytoplasmic surfaces. Urea (8 M) could not remove this fuzzy layer. In electron micrographs of rat ventricular myocytes, cytoplasmic surfaces of gap junctions were fuzzy. We conclude that rat heart gap junction protein consists of an intramembrane component (Mr 29,500) that extends into the “gap” and a cytoplasmic surface component (Mr 14,500–17,500) that corresponds to the fuzzy layer and is hydrolyzable by a serine protease.

scholarly journals Proteinaceous Protease Inhibitor from Lawsonia Inermis: Purification, Characterization and Antibacterial Activity

2013 ◽  
Vol 8 (10) ◽  
pp. 1934578X1300801 ◽  
Author(s):  
Arvind Dabhade ◽  
Priti Patel ◽  
Ulhas Patil
Keyword(s):  
Antibacterial Activity ◽  
Protease Inhibitor ◽  
Deae Cellulose ◽  
Gel Permeation Chromatography ◽  
Apparent Molecular Weight ◽  
Exchange Chromatography ◽  
Lawsonia Inermis ◽  
Sds Page ◽  
Wide Range ◽  
And Staphylococcus Aureus

A thermo-stable, proteinaceous protease inhibitor (LPI) from Lawsonia inermis is reported. The LPI was purified from Lawsonia inermis seeds by subsequent ammonium sulfate precipitation, ion exchange chromatography (DEAE-Cellulose) and gel permeation chromatography (Sephadex-50). The purified protease inhibitor is effective against a wide range of proteases viz. papain, trypsin, pepsin and metallo-protease. The apparent molecular weight of the protease inhibitor is 19 kDa, determined by SDS-PAGE electrophoresis. The protease inhibitor was found to be stable at 70 °C for 30 min. It was also examined for antibacterial activity against Pseudomonas aeruginosa MTCC 7926 and Staphylococcus aureus NCIM 2079; the IC50 values of the purified LPI were 11.4 μg/mL and 16.6 μg/mL respectively.

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