scholarly journals Loss of activity mutations in phospholipase C zeta (PLC ) abolishes calcium oscillatory ability of human recombinant protein in mouse oocytes

2011 ◽  
Vol 26 (12) ◽  
pp. 3372-3387 ◽  
Author(s):  
J. Kashir ◽  
C. Jones ◽  
H. C. Lee ◽  
K. Rietdorf ◽  
D. Nikiforaki ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Phospholipase C ◽  
Mouse Oocytes ◽  
Human Recombinant Protein

scholarly journals Preparation and Characterization of Human Recombinant Protein 1/Clara Cell M r 10 000 Protein

1996 ◽  
Vol 34 (9) ◽  
Author(s):  
Ryuta Okutani ◽  
Yoshihisa Itoh ◽  
Toshiyuki Yamada ◽  
Tetsuji Yamaguchi ◽  
Gurmukh Singh ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Clara Cell ◽  
Human Recombinant Protein

scholarly journals Phospholipase C in mouse oocytes: characterization of β and γ isoforms and their possible involvement in sperm-induced Ca2+ spiking

1996 ◽  
Vol 316 (2) ◽  
pp. 583-591 ◽  
Author(s):  
Genevieve DUPONT ◽  
Orla M. McGUINNESS ◽  
Martin H. JOHNSON ◽  
Michael J. BERRIDGE ◽  
Franck BORGESE
Keyword(s):  
Monoclonal Antibodies ◽  
Phospholipase C ◽  
Kinase Inhibitors ◽  
Critical Role ◽  
Inhibitory Effect ◽  
Functional Studies ◽  
Mouse Oocytes ◽  
Mature Mouse

This study involved an investigation of the role of phospholipase C (PLC) in generating repetitive Ca2+ spikes at fertilization. Using a PCR-based strategy we have demonstrated that mouse oocytes have mRNA coding for PLCβ1, PLCβ3 and PLCγ isoenzymes. Furthermore, immunodetection of PLCγ1 using monoclonal antibodies reveals that PLCγ1 protein is present in mature mouse oocytes, ruling out the possibility that mRNA was being transcribed but not expressed. We were unsuccessful at detecting the presence of PLCβ protein, but the presence of this isoform can be inferred from functional studies. The PLC inhibitor, U73122, exerted an inhibitory effect on oocytes activated by spermatozoa or acetylcholine at concentrations of 10 and 30 μM respectively, while its inactive analogue had no effect. The soluble tyrosine kinase inhibitors, genistein (100 μM), herbimycin (10 μM) and geldanamycin (0.6 μM) which could affect signalling through PLCγ hindered but never completely inhibited Ca2+ spiking in response to fertilization. We conclude that the activation of PLC to generate InsP3 may play a critical role in fertilization.

scholarly journals Plasmodium falciparum Phospholipase C Hydrolyzing Sphingomyelin and Lysocholinephospholipids Is a Possible Target for Malaria Chemotherapy

2001 ◽  
Vol 195 (1) ◽  
pp. 23-34 ◽  
Author(s):  
Kentaro Hanada ◽  
Nirianne Marie Q. Palacpac ◽  
Pamela A. Magistrado ◽  
Ken Kurokawa ◽  
Ganesh Rai ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Recombinant Protein ◽  
Phospholipase C ◽  
Dependent Manner ◽  
Glutathione S Transferase ◽  
Trophozoite Stage ◽  
Severe Impairment ◽  
Active Transcription ◽  
Biochemical Analyses ◽  
Dose Dependent

Sphingomyelinase (SMase) is one of the principal enzymes in sphingomyelin (SM) metabolism. Here, we identified a Plasmodium falciparum gene (PfNSM) encoding a 46-kD protein, the amino acid sequence of which is ∼25% identical to that of bacteria SMases. Biochemical analyses of the recombinant protein GST-PfNSM, a fusion protein of the PfNSM product with glutathione-S-transferase, reveal that this enzyme retained similar characteristics in various aspects to SMase detected in P. falciparum–infected erythrocytes and isolated parasites. In addition, the recombinant protein retains hydrolyzing activity not only of SM but also of lysocholinephospholipids (LCPL) including lysophosphatidylcholine and lysoplatelet-activating factor, indicating that PfNSM encodes SM/LCPL-phospholipase C (PLC). Scyphostatin inhibited SM/LCPL-PLC activities of the PfNSM product as well as the intraerythrocytic proliferation of P. falciparum in a dose-dependent manner with ID50 values for SM/LCPL-PLC activities and the parasite growth at 3–5 μM and ∼7 μM, respectively. Morphological analysis demonstrated most severe impairment in the intraerythrocytic development with the addition of scyphostatin at trophozoite stage than at ring or schizont stages, suggesting its effect specifically on the stage progression from trophozoite to schizont, coinciding with the active transcription of PfNSM gene.

scholarly journals Notum Promotes Proliferation and Migration of Oral Squamous Cell Carcinoma Via Shh/p-GSK3β/β-Catenin Pathway

2021 ◽  
Author(s):  
Panpan Yang ◽  
Congshan Li ◽  
Qin Zhou ◽  
Xiaoqi Zhang ◽  
Yuying Kou ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Oral Squamous Cell Carcinoma ◽  
Recombinant Protein ◽  
Western Blot ◽  
Cell Carcinoma ◽  
Bioinformatics Analysis ◽  
Rt Pcr ◽  
And Migration ◽  
Human Recombinant Protein ◽  
Proliferation And Migration

Abstract Background: Oral squamous cell carcinoma (OSCC) is a common malignant tumor of the head and neck, accounting for more than 90% of oral and maxillofacial malignancies. Therefore, it is of great importance to explore the key factors regulating OSCC. Notum belongs to the α/β hydrolase family and is a deacylated extracellular protein that regulates Wnt and other signaling pathways. Studies have found that Notum participates in the progression of colorectal cancer and hepatocellular carcinoma and targeting Notum can regulate the occurrence of cancer. However, the relationship between Notum and OSCC is currently unclear. This study aims to explore the role of Notum in regulating OSCC and its specific mechanism. Methods: Nine OSCC tissue sections were selected for hematoxylin and eosin staining and immunohistochemistry for PCNA and Notum. Bioinformatics analysis was used to explore the correlation between OSCC and Notum expression. CCK8, Western blot, RT-PCR, scratch experiment, transwell, clone formation, flow cytometry, cellular immunofluorescence and in vivo nude mouse tumor formation methods were applied to OSCC cells treated with Notum human recombinant protein, a Notum overexpression plasmid, and a Notum small interfering RNA to explore the mechanism of Notum in regulating OSCC. Results: The results of immunohistochemistry and bioinformatics analysis showed that Notum was highly expressed in OSCC tissues. Notum human recombinant protein promoted the proliferation and migration of Cal-27 and SCC-15 cells; overexpression of Notum promoted the proliferation and migration of Cal-27 and SCC-15 cells; si-Notum can significantly inhibit proliferation and migration of Cal-27 and SCC-15 cells and promote their apoptosis. Western blot and RT-PCR results showed that si-Notum can inhibit the Hedgehog signaling pathway. In addition, Notum human recombinant protein can increase the phosphorylation level of GSK3β and promote the expression of β-catenin, thereby further regulating the biological behavior of OSCC.Conclusions: Notum regulates the proliferation and migration of OSCC through Shh/p-GSK3β/β-catenin signaling pathway. Notum maybe an effective targets for the treatment of OSCC.

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