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60S acidic ribosomal protein P2

A Bautista-Santos, S Zinker
The four structural acidic ribosomal proteins that dissociate from P1A/P2B and P1B/P2A heterodimers of Saccharomyces cerevisiae were searched in the 60S ribosomal subunit, the 80S monosome, and the polysomal fractions after ribosome profile centrifugation in sucrose gradients in TMN buffer, and after dissociation of monosomes and polysomes to small and large ribosomal subunits in LMS buffer. Analysis by isoelectric focusing, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and Western blotting of these fractions or the purified acidic protein samples showed eight bands that correspond to the acidic ribosomal proteins in the 60S dissociated subunits of the 80S monosome and polysomes...
July 1, 2014: Biochemistry
Antonio Jiménez-Díaz, Miguel Remacha, Juan P G Ballesta, Juan José Berlanga
Eukaryotic cells contain an unusually large cytoplasmic pool of P1/P2 phosphoproteins, which form the highly flexible 60S subunit stalk that is required to interact with and activate soluble translation factors. In cells, cytoplasmic P1/P2 proteins are exchanged for ribosome-bound proteins in a process that can modulate ribosome function and translation. Here, we analysed different S. cerevisiae stalk mutants grown under stress conditions that result in eIF2α phosphorylation. These mutants either lack a cytoplasmic pool of stalk proteins or contain free but not ribosome-bound proteins...
2013: PloS One
Sudipta Das, Himanish Basu, Reshma Korde, Rita Tewari, Shobhona Sharma
Malaria parasites reside inside erythrocytes and the disease manifestations are linked to the growth inside infected erythrocytes (IE). The growth of the parasite is mostly confined to the trophozoite stage during which nuclear division occurs followed by the formation of cell bodies (schizogony). The mechanism and regulation of schizogony are poorly understood. Here we show a novel role for a Plasmodium falciparum 60S stalk ribosomal acidic protein P2 (PfP2) (PFC0400w), which gets exported to the IE surface for 6-8 hrs during early schizogony, starting around 26-28 hrs post-merozoite invasion...
2012: PLoS Pathogens
Pushpa Mishra, Sudipta Das, Lata Panicker, Madhusoodan V Hosur, Shobhona Sharma, Ramakrishna V Hosur
The eukaryotic 60S-ribosomal stalk is composed of acidic ribosomal proteins (P1 and P2) and neutral protein P0, which are thought to be associated as a pentameric structure, [2P1, 2P2, P0]. Plasmodium falciparum P2 (PfP2) appears to play additional non-ribosomal functions associated with its tendency for homo-oligomerization. Recombinant bacterially expressed PfP2 protein also undergoes self-association, as shown by SDS-PAGE analysis and light scattering studies. Secondary structure prediction algorithms predict the native PfP2 protein to be largely helical and this is corroborated by circular dichroism investigation...
2012: PloS One
David Cárdenas, Jesús Revuelta-Cervantes, Antonio Jiménez-Díaz, Hendricka Camargo, Miguel Remacha, Juan P G Ballesta
The ribosomal stalk is formed by four acidic phosphoproteins in Saccharomyces cerevisiae, P1α, P1β, P2α and P2β, which form two heterodimers, P1α/P2β and P1β/P2α, that preferentially bind to sites A and B of the P0 protein, respectively. Using mutant strains carrying only one of the four possible P1/P2 combinations, we found a specific phenotype associated to each P1/P2 pair, indicating that not all acidic P proteins play the same role. The absence of one P1/P2 heterodimer reduced the rate of cell growth by varying degrees, depending on the proteins missing...
May 2012: Nucleic Acids Research
Jeffrey W Priest, James P Kwon, Joel M Montgomery, Caryn Bern, Delynn M Moss, Amanda R Freeman, Cara C Jones, Michael J Arrowood, Kimberly Y Won, Patrick J Lammie, Robert H Gilman, Jan R Mead
Cryptosporidium infection is commonly observed among children and immunocompromised individuals in developing countries, but large-scale outbreaks of disease among adults have not been reported. In contrast, outbreaks of cryptosporidiosis in the United States and Canada are increasingly common among patients of all ages. Thus, it seems likely that residents of regions where Cryptosporidium is highly endemic acquire some level of immunity, while residents of the developed world do not. A new immunodominant Cryptosporidium parvum antigen in the 15- to 17-kDa size range was identified as the Cryptosporidium parvum 60S acidic ribosomal protein P2 (CpP2)...
June 2010: Clinical and Vaccine Immunology: CVI
Peng Zhao, Lijuan Zhang, Weijian Zhong, Xianping Ying, He Huang, Biyun Yao, Zhun Yuan, Juanling Fu, Yuedan Wang, Zongcan Zhou
In the present study, we developed a novel proteomic research strategy named antigen-subtracted 2-DE/MS strategy and applied it to comparative proteomic analysis of anti-benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-transformed human bronchial epithelial cell line (16HBE-C) and its parental cell line (16HBE) G0/G1 cells. Following pre-purification by ammonium sulfate precipitation, rabbit antibodies against 16HBE G0/G1 cells were coupled with protein A/G PLUS-agarose under the maximal coupling rate of about 50%...
January 2011: Archives of Toxicology
Dah-Won Gin, Hye-Rin Kim, Dong-Min Lee, Seung-Hun Lee, Ah-Reum Seong, Ju-Ae Hwang, Joo-Hung Park
When pregnant mice were exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), the average time to eye opening in the offspring was shortened by about a day. How acceleration of eye opening by TCDD occurs remains unknown. To reveal the underlying mechanisms of the accelerated eye opening, pregnant mice were intraperitoneally injected with corn oil or TCDD at GD (gestation day) 11, and tissues around the eye of neonatal mice were subject to proteome analysis and RT-PCR. Upon TCDD administration, translationally controlled tumor protein (TCTP) and 60S acidic ribosomal protein p2 (RLA2) were reduced, while stathmin 1(STMN1) was increased, at both protein and mRNA levels...
July 1, 2010: Toxicology Letters
Ing-Feng Chang, Kathleen Szick-Miranda, Songqin Pan, Julia Bailey-Serres
Analysis of 80S ribosomes of Arabidopsis (Arabidopsis thaliana) by use of high-speed centrifugation, sucrose gradient fractionation, one- and two-dimensional gel electrophoresis, liquid chromatography purification, and mass spectrometry (matrix-assisted laser desorption/ionization time-of-flight and electrospray ionization) identified 74 ribosomal proteins (r-proteins), of which 73 are orthologs of rat r-proteins and one is the plant-specific r-protein P3. Thirty small (40S) subunit and 44 large (60S) subunit r-proteins were confirmed...
March 2005: Plant Physiology
Francesco Giorgianni, Sarka Beranova-Giorgianni, Dominic M Desiderio
Post-translational modifications of proteins from the human pituitary gland play an important role in the regulation of different body functions. We report on the application of a liquid chromatography-tandem mass spectrometry (MS/MS) based approach to detect and characterize phosphorylated proteins in a whole human pituitary digest. By combining an immobilized metal affinity column-based enrichment method with MS/MS conditions that favor the neutral loss of phosphoric acid from a phosphorylated precursor ion, we identified several previously undescribed phosphorylated peptides...
March 2004: Proteomics
Michael Hoff, Ralph M Trüeb, Barbara K Ballmer-Weber, Stefan Vieths, Brunello Wuethrich
BACKGROUND: Quorn is the brand name for a line of foods made with so-called "mycoprotein," which springs from the mold Fusarium venenatum. Since the introduction on the food market, there have been complaints from consumers reporting adverse gastrointestinal reactions after ingestion of mycoprotein. To date, it is not clear whether the reported symptoms are IgE-mediated. OBJECTIVE: The aim of the study was to describe for the first time a case history of an asthmatic patient with severe hypersensitivity reactions to ingested mycoprotein and to identify and characterize the potential allergen that might be responsible for this...
May 2003: Journal of Allergy and Clinical Immunology
M Hoff, B K Ballmer-Weber, B Niggemann, A Cistero-Bahima, M San Miguel-Moncín, A Conti, D Haustein, S Vieths
BACKGROUND: High quality and stability are essential requirements of commercial allergen preparations. Recently we have demonstrated the very low stability of protein allergens in an extract of the ubiquitous mould Fusarium culmorum. OBJECTIVE: The present study was performed to identify, isolate and characterise allergens of F. culmorum as a basis for a stable allergenic reference material. In addition, the significance of IgE binding to carbohydrate structures in the natural allergen source was investigated...
May 2003: Molecular Immunology
Jennifer L R Freeman, Philippe Gonzalo, Julie A Pitcher, Audrey Claing, Jean-Pierre Lavergne, Jean-Paul Reboud, Robert J Lefkowitz
G protein-coupled receptor kinases are well characterized for their ability to phosphorylate and desensitize G protein-coupled receptors (GPCRs). In addition to phosphorylating the beta2-adrenergic receptor (beta2AR) and other receptors, G protein-coupled receptor kinase 2 (GRK2) can also phosphorylate tubulin, a nonreceptor substrate. To identify novel nonreceptor substrates of GRK2, we used two-dimensional gel electrophoresis to find cellular proteins that were phosphorylated upon agonist-stimulation of the beta2AR in a GRK2-dependent manner...
October 22, 2002: Biochemistry
Marek Tchórzewski
The acidic ribosomal P proteins (pI 3-4) are unique among ribosomal constituents: the only molecules on the ribosomes existing in multiple copies, they form a hetero-oligomeric complex (P1/P2)(2) recognizable as a lateral protuberance on the 60S ribosomal subunit, which is thought to be directly involved in interactions with elongation factors during the course of protein synthesis. The role of P proteins in translation is still vague; however, they might possess two functional roles-the proteins may increase performance of ribosomes and/or change ribosomal specificity toward different subset of mRNAs...
August 2002: International Journal of Biochemistry & Cell Biology
Alon Chen, Ella Kaganovsky, Shai Rahimipour, Nurit Ben-Aroya, Eli Okon, Yitzhak Koch
Gonadotropin-releasing hormone (GnRH) analogues are used for the treatment of breast, prostate, and ovarian cancer. These analogues exert their antitumor effects indirectly by inhibiting the pituitary-gonadal axis, as well as by direct inhibition of proliferation of human breast cancer cells. However, the molecular mechanisms mediating these direct antiproliferative effects are not fully understood. We found that normal and malignant human breast tissue express two forms of the neuropeptide GnRH. Quantitative reverse transcription-PCR shows that mRNA encoding the GnRH-I and GnRH-II peptides are overexpressed in cancerous versus normal tissues obtained from the same patients...
February 15, 2002: Cancer Research
Toshiro Maekawa, Michio Fujihara, Kenzo Ohtsuki
Lactoferricin (LFcin) hydrolyzed from lactoferrin (LF), a major 80 kDa iron-binding protein in milk and other exocrine secretions, was characterized as a potent activator of protein kinase CK2 (CK2) in vitro. Human LFcin (hLFcin) at 0.5 microg stimulated approx. 5-fold CK2 activity [phosphorylation of 60S acidic ribosomal proteins (P0, P1, P2) and Hsp90 (p98)] in a manner similar to other functional proteins with oligo-Arg clusters, such as salmine A1, sperm histone H2B and HIV-1 Rev. Interestingly, this stimulatory effect of hLFcin was significantly reduced when it was phosphorylated by A-kinase in vitro...
January 2002: Biological & Pharmaceutical Bulletin
M Pilecki, A Grzyb, P Zień, O Sekuła, R Szyszka
A protein phosphatase dephosphorylating acidic ribosomal proteins was purified from Saccharomyces cerevisiae ribosome-free extract. It was shown that phosphoproteins from both P1 and P2 subfamilies as well as 60S "core" P0 protein were substrates for the enzyme. The phosphatase can dephosphorylate ribosomes as well as histones and casein but the two last substrates with significantly lower efficiency. It was found that the enzyme activity is Mn(2+)-dependent and inhibited by okadaic acid, tautomycin, cantharidin and nodularin at concentrations typical for protein phosphatase type 2A...
2000: Journal of Basic Microbiology
T Maekawa, S Kosuge, A Karino, T Okano, J Ito, H Munakata, K Ohtsuki
The three casein kinase II (CK-II) phosphate acceptors (p35, p17 and p15) in the Superdex CK-II fraction prepared from a 1.5 M NaCl extract of porcine liver were selectively purified by glycyrrhizin (GL)-affinity column chromatography (HPLC) as a heterocomplex associated with CK-II. Determination of the N-terminal amino acid sequences and immunological tests confirmed that these three CK-II phosphate acceptors belong to the family of 60S acidic ribosomal proteins (P0, P1 and P2). Three polyphenol-containing anti-oxidant compounds [catechin, epigallocatechin gallate (EGCG) and quercetin] inhibited CK-II activity (phosphorylation of these ribosomal P proteins) in a dose-dependent manner in vitro...
January 2000: Biological & Pharmaceutical Bulletin
M Tchórzewski, A Boguszewska, D Abramczyk, N Grankowski
The 60S ribosomal subunits from Saccharomyces cerevisiae contain a set of four acidic proteins named YP1alpha, YP1beta, YP2alpha, and YP2beta. The genes for each were PCR amplified from a yeast cDNA library, sequenced, and expressed in Escherichia coli cells using two expression systems. The first system, pLM1, was used for YP1beta, YP2alpha, and YP2beta. The second one, pT7-7, was used for YP1alpha. Expression in both cases was under the control of a strong inducible T7 promoter. The amount of induced recombinant proteins in the host cells was around 10 to 20% of the total soluble bacterial proteins...
February 1999: Protein Expression and Purification
J Bailey-Serres, S Vangala, K Szick, C H Lee
We determined that ribosomes of seedling roots of maize (Zea mays L.) contain the acidic phosphoproteins (P-proteins) known to form a flexible lateral stalk structure of the 60S subunit of eukaryotic ribosomes. The P-protein stalk, composed of P0, P1, and P2, interacts with elongation factors, mRNA, and tRNA during translation. Acidic proteins of 13 to 15.5 kD were released as a complex from ribosomes with 0.4 M NH4Cl/50% ethanol. Protein and cDNA sequence analysis confirmed that maize ribosomes contain one type of P1, two types of P2, and a fourth and novel P1/P2-type protein...
August 1997: Plant Physiology
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