Product References
Protein Extract Preparation and Co-immunoprecipitation from Caenorhabditis elegans.
Journal of visualized experiments : JoVE
Li L,Zinovyeva AY
Co-immunoprecipitation methods are frequently used to study protein-protein interactions. Confirmation of hypothesized protein-protein interactions or identification of new ones can provide invaluable information about the function of a protein of interest. Some of the traditional methods for extract preparation frequently require labor-intensive and time-consuming techniques. Here, a modified extract preparation protocol using a bead mill homogenizer and metal beads is described as a rapid alte
Sat May 23 00:00:00 UTC 2020
Nuclear targeted Saccharomyces cerevisiae asparagine synthetases associate with the mitotic spindle regardless of their enzymatic activity.
PloS one
Noree C,Sirinonthanawech N
Recently, human asparagine synthetase has been found to be associated with the mitotic spindle. However, this event cannot be seen in yeast because yeast takes a different cell division process via closed mitosis (there is no nuclear envelope breakdown to allow the association between any cytosolic enzyme and mitotic spindle). To find out if yeast asparagine synthetase can also (but hiddenly) have this feature, the coding sequences of green fluorescent protein (GFP) and nuclear localization sign
Mon Feb 01 00:00:00 UTC 2021
Arabidopsis TSO1 and MYB3R1 form a regulatory module to coordinate cell proliferation with differentiation in shoot and root.
Proceedings of the National Academy of Sciences of the United States of America
Wang W,Sijacic P,Xu P,Lian H,Liu Z
Fundamental to plant and animal development is the regulated balance between cell proliferation and differentiation a process intimately tied to cell cycle regulation. In mutations in TSO1 whose animal homolog is LIN54 resulted in severe developmental abnormalities both in shoot and root including shoot meristem fasciation and reduced root meristematic zone. The molecular mechanism that could explain the mutant phenotype is absent. Through a genetic screen we identified 32 suppressors that map
Tue Mar 27 00:00:00 UTC 2018
Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling.
The Journal of cell biology
Verweij FJ,Bebelman MP,Jimenez CR,Garcia-Vallejo JJ,Janssen H,Neefjes J,Knol JC,de Goeij-de Haas R,Piersma SR,Baglio SR,Verhage M,Middeldorp JM,Zomer A,van Rheenen J,Coppolino MG,Hurbain I,Raposo G,Smit MJ,Toonen RFG,van Niel G,Pegtel DM
Exosomes are small endosome-derived extracellular vesicles implicated in cell-cell communication and are secreted by living cells when multivesicular bodies (MVBs) fuse with the plasma membrane (PM). Current techniques to study exosome physiology are based on isolation procedures after secretion precluding direct and dynamic insight into the mechanics of exosome biogenesis and the regulation of their release. In this study we propose real-time visualization of MVB-PM fusion to overcome these lim
Mon Mar 05 00:00:00 UTC 2018
Host Factor SPCS1 Regulates the Replication of Japanese Encephalitis Virus through Interactions with Transmembrane Domains of NS2B.
Journal of virology
Ma L,Li F,Zhang JW,Li W,Zhao DM,Wang H,Hua RH,Bu ZG
Signal peptidase complex subunit 1 (SPCS1) is a newly identified host factor that regulates flavivirus replication but the molecular mechanism is not fully understood. Here using Japanese encephalitis virus (JEV) as a model we investigated the mechanism through which the host factor SPCS1 regulates the replication of flaviviruses. We first validated the regulatory function of SPCS1 in JEV propagation by knocking down and knocking out endogenous SPCS1. The loss of SPCS1 function markedly reduced
Fri Jun 15 00:00:00 UTC 2018
Extramitochondrial Assembly of Mitochondrial Targeting Signal Disrupted Mitochondrial Enzyme Aldehyde Dehydrogenase.
Scientific reports
Noree C
Supramolecular assembly of metabolic enzymes has been studied both in vivo and in vitro for nearly a decade. Experimental evidence has suggested a close relationship between enzymatic activity and enzyme assembly/disassembly. However most cases were studied with the cytosolic enzymes. Here I report the evidence for a mitochondrial enzyme with its ability in forming visible intracellular structures. By removing the mitochondrial targeting sequence yeast mitochondrial enzyme aldehyde dehydrogenase
Wed Apr 18 00:00:00 UTC 2018
Engineering Mycobacteria for the Production of Self-Assembling Biopolyesters Displaying Mycobacterial Antigens for Use as a Tuberculosis Vaccine.
Applied and environmental microbiology
Lee JW,Parlane NA,Rehm BHA,Buddle BM,Heiser A
Tuberculosis (TB) is a disease caused by Mycobacterium tuberculosis or Mycobacterium bovis and still remains one of the world's biggest global health burdens. Recently, engineered polyhydroxyalkanoate (PHA) biobeads produced in both E. coli and Lactococcus lactis displaying mycobacterial antigens were found to induce significant cell mediated immune responses in mice. We observed that such PHA beads contained host cell proteins as impurities which we hypothesized to have the potential to in
Wed Mar 01 00:00:00 UTC 2017
Olfactomedin 1 negatively regulates NF-κB signalling and suppresses the growth and metastasis of colorectal cancer cells.
The Journal of pathology
Shi W,Ye Z,Zhuang L,Li Y,Shuai W,Zuo Z,Mao X,Liu R,Wu J,Chen S,Huang W
Uncontrolled growth and distant metastasis are hallmarks of colorectal cancer (CRC), but the mechanisms are poorly understood. Olfactomedin 1 (OLFM1), a member of the olfactomedin domain-containing protein family, plays an important role in the development of neurogenic tissues. Recently, OLFM1 deregulation was frequently observed in several cancers, and it was induced in colon cell lines after treatment with the demethylating agent 5-aza-2'-deoxycytidine. However, the function of OLFM1 in
Tue Nov 01 00:00:00 UTC 2016
The Kaposi's sarcoma-associated herpesvirus ORF34 protein binds to HIF-1α and causes its degradation via the proteasome pathway.
Journal of virology
Haque M,Kousoulas KG
Kaposi's sarcoma-associated herpesvirus (KSHV) is the causative agent for Kaposi's sarcoma (KS) and two other lymphoproliferative disorders, primary effusion lymphoma (PEL) and multicentric Castleman's disease (MCD). Kaposi's sarcoma is a highly vascular tumor, and recently both hypoxia-inducible factor 1α (HIF-1α) and HIF-2α were detected in KS samples, indicating a role of HIFs in the KSHV life cycle. Previously, we showed that ORF34, a lytic gene of unass
Fri Feb 01 00:00:00 UTC 2013
Oncogenic K-ras segregates at spatially distinct plasma membrane signaling platforms according to its phosphorylation status.
Journal of cell science
Barceló C,Paco N,Beckett AJ,Alvarez-Moya B,Garrido E,Gelabert M,Tebar F,Jaumot M,Prior I,Agell N
Activating mutations in the K-Ras small GTPase are extensively found in human tumors. Although these mutations induce the generation of a constitutively GTP-loaded, active form of K-Ras, phosphorylation at Ser181 within the C-terminal hypervariable region can modulate oncogenic K-Ras function without affecting the in vitro affinity for its effector Raf-1. In striking contrast, K-Ras phosphorylated at Ser181 shows increased interaction in cells with the active form of Raf-1 and with p110α,
Tue Oct 15 00:00:00 UTC 2013
Nudel/NudE and Lis1 promote dynein and dynactin interaction in the context of spindle morphogenesis.
Molecular biology of the cell
Wang S,Ketcham SA,Schön A,Goodman B,Wang Y,Yates J,Freire E,Schroer TA,Zheng Y
Lis1, Nudel/NudE, and dynactin are regulators of cytoplasmic dynein, a minus end-directed, microtubule-based motor required for proper spindle assembly and orientation. In vitro studies have shown that dynactin promotes processive movement of dynein on microtubules whereas Lis1 causes dynein to enter a persistent force-generating state (referred to here as dynein stall). Yet, how the activities of Lis1, Nudel/NudE, and dynactin are coordinated to regulate dynein remains poorly understood in vivo
Fri Nov 01 00:00:00 UTC 2013
