Anti-MRPL20 Antibody

Total Page：16

File Type：pdf, Size：1020Kb

D225249 Anti-MRPL20 antibody Cat. No. D225249 Package 25 μl/100 μl/200 μl Storage -20°C, pH7.4 PBS, 0.05% NaN3, 40% Glycerol Product overview Description A n ti-MRPL20 rabbit polyclonal antibody Applications ELISA, WB, IHC Immunogen Fusion protein of human MRPL20 Reactivity Human, Mouse Content 1 mg/ml Host species Rabbit Ig class Immunogen-specific rabbit IgG Purification Antigen affinity purification Target information Symbol MRPL20 Full name mitochondrial ribosomal protein L20 Synonyms L20mt; MRP-L20 Swissprot Q9BYC9 Target Background Mammalian mitochondrial ribosomal proteins are encoded by nuclear genes and help in protein synthesis within the mitochondrion. Mitochondrial ribosomes (mitoribosomes) consist of a small 28S subunit and a large 39S subunit. They have an estimated 75% protein to rRNA composition compared to prokaryotic ribosomes, where this ratio is reversed. Another difference between mammalian mitoribosomes and prokaryotic ribosomes is that the latter contain a 5S rRNA. Among different species, the proteins comprising the mitoribosome differ greatly in sequence, and sometimes in biochemical properties, which prevents easy recognition by sequence homology. This gene encodes a 39S subunit protein. A pseudogene corresponding to this gene is found on chromosome 21q. Alternative splicing results in multiple transcript variantsS encodingangon different isoforms. Biotech Applications Immunohistochemistry Predicted cell location: Cytoplasm Predicted cell location: Cytoplasm Positive control: Human liver cancer Positive control: Human brain Recommended dilution: 50-200 Recommended dilution: 50-200 The image on the left is immunohistochemistry of paraffin-embedded The image on the left is immunohistochemistry of paraffin-embedded Human liver cancer tissue using D225249(MRPL20 Antibody) at Human brain tissue using D225249(MRPL20 Antibody) at dilution dilution 1/60, on the right is treated with fusion protein. (Original 1/60, on the right is treated with fusion protein. (Original magnification: ×200) magnification: ×200) Western blotting Predicted band size:17 kDa Positive control:RAW264.7 cell lysate Recommended dilution: 500-2000 Gel: 12%SDS-PAGE Lysate: 40 μg Lane: RAW264.7 cell lysate Primary antibody: D225249(MRPL20 Antibody) at dilution 1/900 Secondary antibody: Goat anti rabbit IgG at 1/5000 dilution Exposure time: 3 minutes ELISA Sangon Biotech Recommended dilution: 5000-10000 .

Copy Link

Recommended publications

Anti-MRPS17 Monoclonal Antibody, Clone FQS23694 (DCABH-6110) This Product Is for Research Use Only and Is Not Intended for Diagnostic Use
Anti-MRPS17 monoclonal antibody, clone FQS23694 (DCABH-6110) This product is for research use only and is not intended for diagnostic use. PRODUCT INFORMATION Product Overview Rabbit monoclonal to MRPS17 Antigen Description Mammalian mitochondrial ribosomal proteins are encoded by nuclear genes and help in protein synthesis within the mitochondrion. Mitochondrial ribosomes (mitoribosomes) consist of a small 28S subunit and a large 39S subunit. They have an estimated 75% protein to rRNA composition compared to prokaryotic ribosomes, where this ratio is reversed. Another difference between mammalian mitoribosomes and prokaryotic ribosomes is that the latter contain a 5S rRNA. Among different species, the proteins comprising the mitoribosome differ greatly in sequence, and sometimes in biochemical properties, which prevents easy recognition by sequence homology. This gene encodes a 28S subunit protein that belongs to the ribosomal protein S17P family. The encoded protein is moderately conserved between human mitochondrial and prokaryotic ribosomal proteins. Pseudogenes corresponding to this gene are found on chromosomes 1p, 3p, 6q, 14p, 18q, and Xq. Immunogen Recombinant fragment within Human MRPS17. The exact sequence is proprietary.Database link: Q9Y2R5 Isotype IgG Source/Host Rabbit Species Reactivity Human Clone FQS23694 Purity Tissue culture supernatant Conjugate Unconjugated Applications IHC-P, WB, ICC/IF, IP Positive Control HeLa, HepG2 and U937 cell lysate; Human kidney and liver tissue; HeLa cells Format Liquid Size 100 μl Buffer pH: 7.2; Preservative: 0.01% Sodium azide; Constituents: 49% PBS, 0.05% BSA, 50% Glycerol 45-1 Ramsey Road, Shirley, NY 11967, USA Email: [email protected] Tel: 1-631-624-4882 Fax: 1-631-938-8221 1 © Creative Diagnostics All Rights Reserved Preservative 0.01% Sodium Azide Storage Store at +4°C short term (1-2 weeks).
[Show full text]
Micrornas Mediated Regulation of the Ribosomal Proteins and Its Consequences on the Global Translation of Proteins
cells Review microRNAs Mediated Regulation of the Ribosomal Proteins and Its Consequences on the Global Translation of Proteins Abu Musa Md Talimur Reza 1,2 and Yu-Guo Yuan 1,3,* 1 Jiangsu Co-Innovation Center of Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; [email protected] 2 Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawi´nskiego5a, 02-106 Warsaw, Poland 3 Jiangsu Key Laboratory of Zoonosis/Joint International Research Laboratory of Agriculture and Agri-Product Safety, The Ministry of Education of China, Yangzhou University, Yangzhou 225009, China * Correspondence: [email protected]; Tel.: +86-514-8797-9228 Abstract: Ribosomal proteins (RPs) are mostly derived from the energy-consuming enzyme families such as ATP-dependent RNA helicases, AAA-ATPases, GTPases and kinases, and are important structural components of the ribosome, which is a supramolecular ribonucleoprotein complex, composed of Ribosomal RNA (rRNA) and RPs, coordinates the translation and synthesis of proteins with the help of transfer RNA (tRNA) and other factors. Not all RPs are indispensable; in other words, the ribosome could be functional and could continue the translation of proteins instead of lacking in some of the RPs. However, the lack of many RPs could result in severe defects in the biogenesis of ribosomes, which could directly inﬂuence the overall translation processes and global expression of the proteins leading to the emergence of different diseases including cancer. While microRNAs (miRNAs) are small non-coding RNAs and one of the potent regulators of the post-transcriptional 0 gene expression, miRNAs regulate gene expression by targeting the 3 untranslated region and/or coding region of the messenger RNAs (mRNAs), and by interacting with the 50 untranslated region, Citation: Reza, A.M.M.T.; Yuan, Y.-G.
[Show full text]
Mitochondrial Translation and Its Impact on Protein Homeostasis And
Mitochondrial translation and its impact on protein homeostasis and aging Tamara Suhm Academic dissertation for the Degree of Doctor of Philosophy in Biochemistry at Stockholm University to be publicly defended on Friday 15 February 2019 at 09.00 in Magnélisalen, Kemiska övningslaboratoriet, Svante Arrhenius väg 16 B. Abstract Besides their famous role as powerhouse of the cell, mitochondria are also involved in many signaling processes and metabolism. Therefore, it is unsurprising that mitochondria are no isolated organelles but are in constant crosstalk with other parts of the cell. Due to the endosymbiotic origin of mitochondria, they still contain their own genome and gene expression machinery. The mitochondrial genome of yeast encodes eight proteins whereof seven are core subunits of the respiratory chain and ATP synthase. These subunits need to be assembled with subunits imported from the cytosol to ensure energy supply of the cell. Hence, coordination, timing and accuracy of mitochondrial gene expression is crucial for cellular energy production and homeostasis. Despite the central role of mitochondrial translation surprisingly little is known about the molecular mechanisms. In this work, I used baker’s yeast Saccharomyces cerevisiae to study different aspects of mitochondrial translation. Exploiting the unique possibility to make directed modifications in the mitochondrial genome of yeast, I established a mitochondrial encoded GFP reporter. This reporter allows monitoring of mitochondrial translation with different detection methods and enables more detailed studies focusing on timing and regulation of mitochondrial translation. Furthermore, employing insights gained from bacterial translation, we showed that mitochondrial translation efficiency directly impacts on protein homeostasis of the cytoplasm and lifespan by affecting stress handling.
[Show full text]
The Role of Human Ribosomal Proteins in the Maturation of Rrna and Ribosome Production
JOBNAME: RNA 14#9 2008 PAGE: 1 OUTPUT: Friday August 8 17:34:50 2008 csh/RNA/164293/rna11320 Downloaded from rnajournal.cshlp.org on September 27, 2021 - Published by Cold Spring Harbor Laboratory Press The role of human ribosomal proteins in the maturation of rRNA and ribosome production SARA ROBLEDO,1,3 RACHEL A. IDOL,1,3 DAN L. CRIMMINS,2 JACK H. LADENSON,2 PHILIP J. MASON,1,4 and MONICA BESSLER1,4 1Department of Internal Medicine, Division of Hematology, Washington University School of Medicine, St. Louis, Missouri 63110, USA 2Department of Pathology and Immunology, Division of Laboratory and Genomic Medicine, Washington University School of Medicine, St. Louis, Missouri 63110, USA ABSTRACT Production of ribosomes is a fundamental process that occurs in all dividing cells. It is a complex process consisting of the coordinated synthesis and assembly of four ribosomal RNAs (rRNA) with about 80 ribosomal proteins (r-proteins) involving more than 150 nonribosomal proteins and other factors. Diamond Blackfan anemia (DBA) is an inherited red cell aplasia caused by mutations in one of several r-proteins. How defects in r-proteins, essential for proliferation in all cells, lead to a human disease with a specific defect in red cell development is unknown. Here, we investigated the role of r-proteins in ribosome biogenesis in order to find out whether those mutated in DBA have any similarities. We depleted HeLa cells using siRNA for several individual r-proteins of the small (RPS6, RPS7, RPS15, RPS16, RPS17, RPS19, RPS24, RPS25, RPS28) or large subunit (RPL5, RPL7, RPL11, RPL14, RPL26, RPL35a) and studied the effect on rRNA processing and ribosome production.
[Show full text]
Scores Tag L1102 L1214 L1232 HOSE1 HOSE2 HS 1
Scores Tag L1102 L1214 L1232 HOSE1 HOSE2 HS 1 CTGGAGGCTG 9.5 8.71 10.67 0 0 229335 1 CTGGAGGCTG 9.5 8.71 10.67 0 0 169350 1 CTGGAGGCTG 9.5 8.71 10.67 0 0 61384 1 CTGGAGGCTG 9.5 8.71 10.67 0 0 105633 1 CTGGAGGCTG 9.5 8.71 10.67 0 0 149152 1 GCAACTGTGA 7.77 8.71 6.67 0 0 169476 1 ATTTGTCCCA 14.68 7.62 5.33 0 0 57301 1 ATTTGTCCCA 14.68 7.62 5.33 0 0 356122 1 GTCGGGCCTC 71.65 39.18 22.67 1.16 0 73769 1 ATTCTCCAGT 35.39 39.18 44.01 85.74 89.13 458218 1 ATTCTCCAGT 35.39 39.18 44.01 85.74 89.13 406300 1 AGGGCTTCCA 56.98 37 69.35 134.4 141.05 458148 1 CTGCTATACG 11.22 14.15 9.34 41.71 38.94 180946 1 TTGGTGAAGG 10.36 18.5 17.34 61.41 49.32 426138 1 GCCGTGTCCG 21.58 9.79 8 54.45 58.84 356666 1 GCCGTGTCCG 21.58 9.79 8 54.45 58.84 380843 1 ACCCACGTCA 0.86 0 1.33 27.81 20.77 298184 1 ACCCACGTCA 0.86 0 1.33 27.81 20.77 400124 1 TCTCCATACC 0.86 1.09 0 23.17 25.09 1 CCCTTGTCCG 0.86 0 0 26.65 20.77 127824 1 CTTCTTGCCC 0 0 1.33 47.5 36.34 347939 1 CTTCTTGCCC 0 0 1.33 47.5 36.34 424220 1 CTGTACTTGT 0.86 0 0 63.72 29.42 75678 1 CCCAACGCGC 0 0 0 83.42 47.59 347939 1 GCAAGAAAGT 0 0 0 26.65 39.81 36977 1 GCAAGAAAGT 0 0 0 26.65 39.81 155376 1 ACACAGCAAG 0 0 0 23.17 15.58 1 AGCTTCCACC 0 0 0 11.59 7.79 355542 1 GAGTGGCTAC 0 0 0 9.27 6.92 1 ATGGTGGGGG 0 0 0 8.11 22.5 343586 1 AGATCCCAAG 0 0 0 5.79 8.65 50813 1 TGGAAGGAGG 0 0 0 8.11 6.06 1 TAGCCGGGAC 0 0 0 5.79 7.79 107740 1 TGTGGATGTG 0 0 0 4.63 12.11 180878 1 GGGTAGGGGG 0 0 0 34.76 9.52 13323 0.99 AATAAAGCAA 2.59 7.62 8 0 0 76698 0.99 AATAAAGCAA 2.59 7.62 8 0 0 126043 0.99 GGAACAAACA 8.63 3.26 18.67 0 0 375108
[Show full text]
RPSA Gene Ribosomal Protein SA
RPSA gene ribosomal protein SA Normal Function The RPSA gene provides instructions for making a protein called ribosomal protein SA, which is one of approximately 80 different ribosomal proteins. These proteins come together to form structures called ribosomes. Ribosomes process the cell's genetic instructions to create proteins. Each ribosome is made up of two parts (subunits) called the large subunit and the small subunit. Ribosomal protein SA is part of the small subunit. The specific roles of each of the ribosomal proteins within the ribosome are not entirely understood. Some ribosomal proteins are involved in the assembly or stability of ribosomes. Others help carry out the ribosome's main function of building new proteins. Research suggests that ribosomal protein SA helps the ribosome control the production of certain proteins, many of which are likely important for development before birth. Health Conditions Related to Genetic Changes Isolated congenital asplenia At least 20 RPSA gene mutations have been identified in individuals with isolated congenital asplenia. People with this condition do not have a spleen but have no other developmental abnormalities. The spleen plays an important role in the immune system. Without this organ, affected individuals are highly susceptible to bacterial infections, which can be life-threatening. RPSA gene mutations are thought to reduce the amount of functional ribosomal protein SA. A shortage of the normal protein likely impairs the assembly of ribosomes, but the specific effects of the mutations
[Show full text]
Elabscience.Com ® E-Mail:[email protected] Elabscience Elabscience Biotechnology Inc
Tel:240-252-7368(USA) Fax:240-252-7376(USA) www.elabscience.com ® E-mail:[email protected] Elabscience Elabscience Biotechnology Inc. MRPL20 Polyclonal Antibody Catalog No. E-AB-18777 Reactivity H,M Storage Store at -20℃. Avoid freeze / thaw cycles. Host Rabbit Applications WB,IHC,ELISA Isotype IgG Note: Centrifuge before opening to ensure complete recovery of vial contents. Images Immunogen Information Immunogen Fusion protein of human MRPL20 Gene Accession BC009515 Swissprot Q9BYC9 Synonyms 39S ribosomal protein L20,mitochondrial,L20mt,MG C4779,MGC74465,Mitochondrial ribosomal protein L20,MRPL 20 Western blot analysis of RAW264.7 Product Information cell lysate using MRPL20 Polyclonal Calculated MW 17 kDa Antibody at dilution of 1:900 Observed MW Refer to figures Buffer PBS with 0.05% NaN3 and 40% Glycerol,pH7.4 Purify Antigen affinity purification Dilution WB 1:500-1:2000, IHC 1:50-1:200, ELISA 1:5000-1:10000 Background MRPL20 is one of more than 70 protein components of mitochondrial Immunohistochemistry of paraffin- ribosomes that are encoded by the nuclear genome. MRPL20 is a subunit embedded Human liver cancer tissue of the 39S mitochondrial ribosome. Mitochondrial ribosomes using MRPL20 Polyclonal Antibody at (mitoribosomes) consist of a small 28S subunit and a large 39S subunit. dilution of 1:60(×200) They have an estimated 75% protein to rRNA composition compared to prokaryotic ribosomes, where this ratio is reversed. Another difference between mammalian mitoribosomes and prokaryotic ribosomes is that the latter contain a 5S rRNA. Among different species, the proteins comprising the mitoribosome differ greatly in sequence, and sometimes in biochemical properties, which prevents easy recognition by sequence homology.
[Show full text]
The C-Terminal Extension Domain of Saccharomyces Cerevisiae Mrpl32, a Homolog of Ribosomal Protein L32, Functions in Trans to Support Mitochondrial Translation
Genes Genet. Syst. (2018) 93, p. 21–24 Function of C-terminal extension of MrpL32 21 The C-terminal extension domain of Saccharomyces cerevisiae MrpL32, a homolog of ribosomal protein L32, functions in trans to support mitochondrial translation Ivo Ngundu Woogeng and Madoka Kitakawa* Department of Biology, Faculty of Science, Kobe University, Rokko 1-1, Kobe, Hyogo 657-8501, Japan (Received 3 June 2017, accepted 10 August 2017; J-STAGE Advance published date: 17 January 2018) Mitochondrial ribosomal protein L32 (MrpL32) of Saccharomyces cerevisiae is homologous to the bacterial L32 ribosomal protein. MrpL32 carries an N-terminal mitochondrion-targeting sequence (MTS) and is about 60 amino acid residues lon- ger at the C-terminus. Adding to its function as a leader sequence, the MTS of MrpL32 has been reported to regulate ribosome biogenesis through its processing by m-AAA protease. However, the function of the C-terminal extension (CE) remains totally unknown. Therefore, we constructed a series of C-terminally truncated mrpl32 (mrpl32ΔC) genes and expressed them in a Δmrpl32 mutant to examine their function. Interestingly, some MrpL32ΔC derivatives exhibited temperature- sensitive (ts) growth on medium with non-fermentable carbon sources. Further- more, the CE domain of MrpL32, expressed separately from MrpL32ΔC, could rescue the ts phenotype of mutants by improving mitochondrial protein synthesis. Key words: C-terminal extension, mitochondrial ribosome, MrpL32 Many protein components of mitochondrial ribosomes (Huff et al., 1993). are homologous to eubacterial ribosomal proteins, but they MrpL32 is unique; its long mitochondrion-targeting tend to be larger than the corresponding bacterial proteins sequence (MTS) (71 aa) is a target of m-AAA prote- (Graack and Wittmann-Liebold, 1998; Gan et al., 2002; ase, and the loss of MrpL32 processing is thoroughly Smits et al., 2007).
[Show full text]
Duodenal Mucosal Mitochondrial Gene Expression Is Associated with Delayed Gastric Emptying in Diabetic Gastroenteropathy
Duodenal mucosal mitochondrial gene expression is associated with delayed gastric emptying in diabetic gastroenteropathy Susrutha Puthanmadhom Narayanan, … , Tamas Ordog, Adil E. Bharucha JCI Insight. 2021;6(2):e143596. https://doi.org/10.1172/jci.insight.143596. Research Article Endocrinology Gastroenterology Graphical abstract Find the latest version: https://jci.me/143596/pdf RESEARCH ARTICLE Duodenal mucosal mitochondrial gene expression is associated with delayed gastric emptying in diabetic gastroenteropathy Susrutha Puthanmadhom Narayanan,1 Daniel O’Brien,2 Mayank Sharma,1 Karl Miller,3 Peter Adams,3 João F. Passos,4 Alfonso Eirin,5 Tamas Ordog,4 and Adil E. Bharucha1 1Division of Gastroenterology and Hepatology, Department of Medicine, Mayo Clinic, Rochester, Minnesota, USA. 2Department of Biomedical Statistics and Informatics, Mayo Clinic, Rochester, Minnesota, USA. 3Sanford Burnham Prebys Medical Discovery Institute, San Diego, California, USA. 4Department of Physiology and Biomedical Engineering and 5Division of Nephrology & Hypertension Research, Department of Medicine, Mayo Clinic, Rochester, Minnesota, USA. Hindered by a limited understanding of the mechanisms responsible for diabetic gastroenteropathy (DGE), management is symptomatic. We investigated the duodenal mucosal expression of protein- coding genes and microRNAs (miRNA) in DGE and related them to clinical features. The diabetic phenotype, gastric emptying, mRNA, and miRNA expression and ultrastructure of duodenal mucosal biopsies were compared in 39 DGE patients and 21 controls. Among 3175 differentially expressed genes (FDR < 0.05), several mitochondrial DNA–encoded (mtDNA-encoded) genes (12 of 13 protein coding genes involved in oxidative phosphorylation [OXPHOS], both rRNAs and 9 of 22 transfer RNAs) were downregulated; conversely, nuclear DNA–encoded (nDNA-encoded) mitochondrial genes (OXPHOS) were upregulated in DGE.
[Show full text]
An Update on Mitochondrial Ribosome Biology: the Plant Mitoribosome in the Spotlight
cells Review An Update on Mitochondrial Ribosome Biology: The Plant Mitoribosome in the Spotlight Artur Tomal y , Malgorzata Kwasniak-Owczarek y and Hanna Janska * Department of Cellular Molecular Biology, Faculty of Biotechnology, University of Wroclaw, 50-383 Wroclaw, Poland; [email protected] (A.T.); [email protected] (M.K.-O.) * Correspondence: [email protected]; Tel.: +0048-713-756-249; Fax: +0048-713-756-234 These authors contributed equally to this work. y Received: 31 October 2019; Accepted: 1 December 2019; Published: 3 December 2019 Abstract: Contrary to the widely held belief that mitochondrial ribosomes (mitoribosomes) are highly similar to bacterial ones, recent experimental evidence reveals that mitoribosomes do differ significantly from their bacterial counterparts. This review is focused on plant mitoribosomes, but we also highlight the most striking similarities and differences between the plant and non-plant mitoribosomes. An analysis of the composition and structure of mitoribosomes in trypanosomes, yeast, mammals and plants uncovers numerous organism-specific features. For the plant mitoribosome, the most striking feature is the enormous size of the small subunit compared to the large one. Apart from the new structural information, possible functional peculiarities of different types of mitoribosomes are also discussed. Studies suggest that the protein composition of mitoribosomes is dynamic, especially during development, giving rise to a heterogeneous populations of ribosomes fulfilling specific functions. Moreover, convincing data shows that mitoribosomes interact with components involved in diverse mitochondrial gene expression steps, forming large expressosome-like structures. Keywords: mitochondrial ribosome; ribosomal proteins; ribosomal rRNA; PPR proteins; translation; plant mitoribosome 1.
[Show full text]
Inhibition of the MID1 Protein Complex
Matthes et al. Cell Death Discovery (2018) 4:4 DOI 10.1038/s41420-017-0003-8 Cell Death Discovery ARTICLE Open Access Inhibition of the MID1 protein complex: a novel approach targeting APP protein synthesis Frank Matthes1,MoritzM.Hettich1, Judith Schilling1, Diana Flores-Dominguez1, Nelli Blank1, Thomas Wiglenda2, Alexander Buntru2,HannaWolf1, Stephanie Weber1,InaVorberg 1, Alina Dagane2, Gunnar Dittmar2,3,ErichWanker2, Dan Ehninger1 and Sybille Krauss1 Abstract Alzheimer’s disease (AD) is characterized by two neuropathological hallmarks: senile plaques, which are composed of amyloid-β (Aβ) peptides, and neurofibrillary tangles, which are composed of hyperphosphorylated tau protein. Aβ peptides are derived from sequential proteolytic cleavage of the amyloid precursor protein (APP). In this study, we identified a so far unknown mode of regulation of APP protein synthesis involving the MID1 protein complex: MID1 binds to and regulates the translation of APP mRNA. The underlying mode of action of MID1 involves the mTOR pathway. Thus, inhibition of the MID1 complex reduces the APP protein level in cultures of primary neurons. Based on this, we used one compound that we discovered previously to interfere with the MID1 complex, metformin, for in vivo experiments. Indeed, long-term treatment with metformin decreased APP protein expression levels and consequently Aβ in an AD mouse model. Importantly, we have initiated the metformin treatment late in life, at a time-point where mice were in an already progressed state of the disease, and could observe an improved behavioral phenotype. These 1234567890 1234567890 findings together with our previous observation, showing that inhibition of the MID1 complex by metformin also decreases tau phosphorylation, make the MID1 complex a particularly interesting drug target for treating AD.
[Show full text]
Interaction of Escherichia Coli Ribosomal Protein S1 with Ribosomes
Proc. Nati. Acad. Sci. USA Vol. 76, No. 3, pp. 1040-1044, March 1979 Biochemistry Interaction of Escherichia coli ribosomal protein S1 with ribosomes (SI polynucleotide binding sites/30S ribosomal subunits/S1-30S binding constants/S1-30S binding stoichiometry/ 16S ribosomal RNA) DAVID E. DRAPER* AND PETER H. VON HIPPEL Institute of Molecular Biology and Department of Chemistry, University of Oregon, Eugene, Oregon 97403 Contributed bt Peter H. von Hippel, November 16,1978 ABSTRACT The binding affinity of Escherichia coli ribo- tein to ribosomes and to ribosomal subunits and report here the somal protein SI for 30S ribosomal particles has been deter- results of some preliminary attempts at a quantitation of the mined by a sucrose gradient band sedimentation technique; the association constant (K) for the binding of one SI protein per Si-ribosome binding interaction. active 30S ribosomal subunit is t2 X 108 M-1. The involvement Three basic questions are addressed: (i) What is the binding of the two polynucleotide binding sites of SI protein (site I affinity of S1 for the 30S (and 50S and 70S) ribosomal compo- binding single-stranded DNA or RNA, and site II binding sin- nents? (ii) Does either S1 polynucleotide binding site contribute gle-stranded RNA only) in the SI-ribosomal interaction have been examined by competition experiments with polynucleo- to the Sl-ribosome interaction, or is this binding entirely at- tides of known affinity for the two sites. We find that site I does tributable to interactions with other proteins of the 30S particle? not contribute to the interaction; site II binding appears to (iii) Is either S1 polynucleotide binding site available for in- provide a major part of the binding free energy, presumably by teraction with mRNA when S1 is bound to the ribosome? We interaction of SI with the 16S rRNA of the 30S particle.
[Show full text]