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Han Zhu, Hua Qin, De-Min Li, Jing Liu, Qiu Zhao
The objective of this study was to investigate the effect of silencing gene protein phosphatase 1H (PPM1H) on malignant phenotype of human pancreatic cancer cell line BxPC-3. In order to explore the function of PPM1H in pancreatic cancer cells, real-time PCR and western blotting were used to detect the expression of PPM1H in different pancreatic cancer cell lines. Human pancreatic cancer cell line BxPC-3 was treated with 10 ng/ml TGF-β1 and 200 ng/ml BMP2 for 72 h, respectively, and the mRNA and protein expression levels of PPM1H and EMT-related markers (E-cadherin, vimentin) were detected by real-time PCR and western blotting, respectively...
November 2016: Oncology Reports
Yasunori Sugiyama, Sho Yamashita, Yuuki Uezato, Yukako Senga, Syouichi Katayama, Naoki Goshima, Yasushi Shigeri, Noriyuki Sueyoshi, Isamu Kameshita
To analyze a variety of protein phosphatases, we developed phosphorylated TandeMBP (P-TandeMBP), in which two different mouse myelin basic protein isoforms were fused in tandem, as a protein phosphatase substrate. P-TandeMBP was prepared efficiently in four steps: (1) phosphorylation of TandeMBP by a protein kinase mixture (Ca(2+)/calmodulin-dependent protein kinase Iδ, casein kinase 1δ, and extracellular signal-regulated kinase 2); (2) precipitation of both P-TandeMBP and protein kinases to remove ATP, Pi, and ADP; (3) acid extraction of P-TandeMBP with HCl to remove protein kinases; and (4) neutralization of the solution that contains P-TandeMBP with Tris...
November 15, 2016: Analytical Biochemistry
Tao Shen, Lan Qin, Xia Lin
Phosphorylation of Smad1/5/8 at the C-terminal SXS motif by BMP type I receptors is one of the most critical events in BMP signaling. Conversely, protein phosphatases that dephosphorylate phospho-Smad1/5/8 can consequently prevent or terminate BMP signaling. PPM1H is an undercharacterized phosphatase in the PPM family. We recently demonstrated that PPM1H can dephosphorylate Smad1 in the cytoplasm and block BMP signaling responses in cellular assays. Here we describe in vitro method showing that PPM1H is a bona fide phosphatase for Smad1/5/8...
2016: Methods in Molecular Biology
Tao Shen, Chuang Sun, Zhengmao Zhang, Ningyi Xu, Xueyan Duan, Xin-Hua Feng, Xia Lin
Bone morphogenetic proteins (BMPs) belong to the TGF-β superfamily of structurally related signaling proteins that regulate a wide array of cellular functions. The key step in BMP signal transduction is the BMP receptor-mediated phosphorylation of transcription factors Smad1, 5, and 8 (collectively Smad1/5/8), which leads to the subsequent activation of BMP-induced gene transcription in the nucleus. In this study, we describe the identification and characterization of PPM1H as a novel cytoplasm-localized Smad1/5/8-specific phosphatase...
June 2014: Cell Research
Si Tuen Lee-Hoeflich, Thinh Q Pham, Don Dowbenko, Xander Munroe, James Lee, Li Li, Wei Zhou, Peter M Haverty, Kanan Pujara, Jeremy Stinson, Sara M Chan, Jeffrey Eastham-Anderson, Ajay Pandita, Somasekar Seshagiri, Klaus P Hoeflich, Gulisa Turashvili, Karen A Gelmon, Samuel A Aparicio, David P Davis, Mark X Sliwkowski, Howard M Stern
The HER2 oncogene is overexpressed or amplified in 20% of breast cancers. HER2-positive cancer historically portends a poor prognosis, but the HER2-targeted therapy trastuzumab mitigates this otherwise ominous distinction. Nevertheless, some patients suffer disease recurrence despite trastuzumab, and metastatic disease remains largely incurable due to innate and acquired resistance. Thus, understanding trastuzumab resistance remains an unmet medical need. Through RNA interference screening, we discovered that knockdown of the serine/threonine phosphatase PPM1H confers trastuzumab resistance via reduction in protein levels of the tumor suppressor p27...
September 2011: Cancer Discovery
Nicola Aceto, Mohamed Bentires-Alj
Lee-Hoeflich and colleagues use RNA interference screening to identify the serine/threonine phosphatase PPM1H as an inhibitor of trastuzumab resistance in vitro. This finding extends the molecular portrait of trastuzumab-resistant cells and provides a rationale when searching for potential therapeutic targets among regulators of PPM1H and/or its substrates.
September 2011: Cancer Discovery
Benjamin M Neale, Sarah Medland, Stephan Ripke, Richard J L Anney, Philip Asherson, Jan Buitelaar, Barbara Franke, Michael Gill, Lindsey Kent, Peter Holmans, Frank Middleton, Anita Thapar, Klaus-Peter Lesch, Stephen V Faraone, Mark Daly, Thuy Trang Nguyen, Helmut Schäfer, Hans-Christoph Steinhausen, Andreas Reif, Tobias J Renner, Marcel Romanos, Jasmin Romanos, Andreas Warnke, Susanne Walitza, Christine Freitag, Jobst Meyer, Haukur Palmason, Aribert Rothenberger, Ziarih Hawi, Joseph Sergeant, Herbert Roeyers, Eric Mick, Joseph Biederman
OBJECTIVE: Although twin and family studies have shown attention-deficit/hyperactivity disorder (ADHD) to be highly heritable, genetic variants influencing the trait at a genome-wide significant level have yet to be identified. Thus additional genomewide association studies (GWAS) are needed. METHOD: We used case-control analyses of 896 cases with DSM-IV ADHD genotyped using the Affymetrix 5.0 array and 2,455 repository controls screened for psychotic and bipolar symptoms genotyped using Affymetrix 6...
September 2010: Journal of the American Academy of Child and Adolescent Psychiatry
Silvia N Kariuki, Beverly S Franek, Akaash A Kumar, Jasmine Arrington, Rachel A Mikolaitis, Tammy O Utset, Meenakshi Jolly, Mary K Crow, Andrew D Skol, Timothy B Niewold
INTRODUCTION: Systemic lupus erythematosus (SLE) is a highly heterogeneous disorder, characterized by differences in autoantibody profile, serum cytokines, and clinical manifestations. SLE-associated autoantibodies and high serum interferon alpha (IFN-α) are important heritable phenotypes in SLE which are correlated with each other, and play a role in disease pathogenesis. These two heritable risk factors are shared between ancestral backgrounds. The aim of the study was to detect genetic factors associated with autoantibody profiles and serum IFN-α in SLE...
2010: Arthritis Research & Therapy
Takeyuki Sugiura, Yoshie Noguchi
Protein phosphatase 2C (PP2C) family is characterized by requirement of metal cation for phosphatase activity. We previously established that PPM1H is a cancer-associated member of the PP2C family. Here we further characterized the phosphatase activity of PPM1H, focusing on its dependence on metal cation. PPM1H possesses the potential to dephosphorylate p-nitrophenyl phosphate (pNPP), casein and phosphopeptides. Interestingly, PPM1H shows the metal preference that is varied depending on the substrate (substrate-dependent metal preference); PPM1H prefers Mn(2+) when pNPP or phosphopeptides is used as a substrate...
June 2009: Biometals: An International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine
Sathyavageeswaran Shreeram, Dmitry V Bulavin
No abstract text is available yet for this article.
February 2008: Cancer Biology & Therapy
Takeyuki Sugiura, Yoshie Noguchi, Kayo Sakurai, Chiharu Hattori
In search for a new anticancer drug target, we explored genes involved in colon adenocarcinoma development through dysregulation of a signal transduction pathway. By using the gene expression profile database, we found protein phosphatase 1H (PPM1H), belonging to the protein phosphatase 2C (PP2C) family, upregulated in colon adenocarcinomas compared with normal colon tissues. RT-PCR analysis verified the elevated level of PPM1H expression in colon cancer cell lines relative to a normal colon cell line. PPM1H encodes a protein with a molecular mass of approximately 50 kDa that resides in the cytoplasm...
February 2008: Cancer Biology & Therapy
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