Reply to Bagni: On BC1 RNA and the fragile X mental retardation protein.
AuthorsIacoangeli A, Rozhdestvensky TS, Dolzhanskaya N, Tournier B, Schütt J, Brosius J, Denman RB, Khandjian EW, Kindler S, Tiedge H,
JournalProc Natl Acad Sci U S A
PubMed ID18511554
'The fragile X mental retardation protein (FMRP), the functional absence of which causes fragile X syndrome, is an RNA-binding protein that has been implicated in the regulation of local protein synthesis at the synapse. The mechanism of FMRP’s interaction with its target mRNAs, however, has remained controversial. In one model, ... More
Characterization of human cytochrome P450s involved in the bioactivation of tri-ortho-cresyl phosphate (ToCP).
'Tri-ortho-cresyl phosphate (ToCP) is a multipurpose organophosphorus compound that is neurotoxic and suspected to be involved in aerotoxic syndrome in humans. It has been reported that not ToCP itself but a metabolite of ToCP, namely, 2-(ortho-cresyl)-4H-1,2,3-benzodioxaphosphoran-2-one (CBDP), may be responsible for this effect as it can irreversibly bind to human ... More
Profiling cholinesterase adduction: a high-throughput prioritization method for organophosphate exposure samples.
AuthorsCarter MD, Crow BS, Pantazides BG, Watson CM, DeCastro BR, Thomas JD, Blake TA, Johnson RC
Journal
PubMed ID23954929
'A high-throughput prioritization method was developed for use with a validated confirmatory method detecting organophosphorus nerve agent exposure by immunomagnetic separation high-performance liquid chromatography tandem mass spectrometry. A ballistic gradient was incorporated into this analytical method to profile unadducted butyrylcholinesterase (BChE) in clinical samples. With Zhang et al.''s Z'' factor ... More
A useful approach to total analysis of RISC-associated RNA.
AuthorsHayashida Y, Nishibu T, Inoue K, Kurokawa T,
JournalBMC Res Notes
PubMed ID19706194
'ABSTRACT: BACKGROUND: Identifying the endogenous RNA induced silencing complex(RISC)-associated RNAs is essential for understanding the cellular regulatory networks by miRNAs. Recently, isolation of RISC-associated mRNAs using antibody was reported, but their method needs a large amount of initial materials. We tried to improve the protocol and constructed an efficient and ... More
Nitration and inactivation of IDO by peroxynitrite.
AuthorsFujigaki H, Saito K, Lin F, Fujigaki S, Takahashi K, Martin BM, Chen CY, Masuda J, Kowalak J, Takikawa O, Seishima M, Markey SP,
JournalJ Immunol
PubMed ID16365430
IDO induction can deplete L-tryptophan in target cells, an effect partially responsible for the antimicrobial activities and antiallogeneic T cell responses of IFN-gamma in human macrophages, dendritic cells, and bone marrow cells. L-tryptophan depletion and NO production are both known to have an antimicrobial effect in macrophages, and the interaction ... More
Protein sorting in the late Golgi of Saccharomyces cerevisiae does not require mannosylated sphingolipids.
AuthorsLisman Q, Pomorski T, Vogelzangs C, Urli-Stam D, de Cocq van Delwijnen W, Holthuis JC,
JournalJ Biol Chem
PubMed ID14583628
Glycosphingolipids are widely viewed as integral components of the Golgi-based machinery by which membrane proteins are targeted to compartments of the endosomal/lysosomal system and to the surface domains of polarized cells. The yeast Saccharomyces cerevisiae creates glycosphingolipids by transferring mannose to the head group of inositol phosphorylceramide (IPC), yielding mannosyl-IPC ... More
Dynamic changes in transcription factor complexes during erythroid differentiation revealed by quantitative proteomics.
AuthorsBrand M, Ranish JA, Kummer NT, Hamilton J, Igarashi K, Francastel C, Chi TH, Crabtree GR, Aebersold R, Groudine M,
JournalNat Struct Mol Biol
PubMed ID14718926
During erythroid differentiation, beta-globin gene expression is regulated by the locus control region (LCR). The transcription factor NF-E2p18/MafK binds within this region and is essential for beta-globin expression in murine erythroleukemia (MEL) cells. Here we use the isotope-coded affinity tag (ICAT) technique of quantitative mass spectrometry to compare proteins interacting ... More
Large T antigen promotes JC virus replication in G2-arrested cells by inducing ATM- and ATR-mediated G2 checkpoint signaling.
AuthorsOrba Y, Suzuki T, Makino Y, Kubota K, Tanaka S, Kimura T, Sawa H,
JournalJ Biol Chem
PubMed ID19903823
Large T antigen (TAg) of the human polyomavirus JC virus (JCV) possesses DNA binding and helicase activities, which, together with various cellular proteins, are required for replication of the viral genome. We now show that JCV-infected cells expressing TAg accumulate in the G(2) phase of the cell cycle as a ... More
pLG72 modulates intracellular D-serine levels through its interaction with D-amino acid oxidase: effect on schizophrenia susceptibility.
AuthorsSacchi S, Bernasconi M, Martineau M, Mothet JP, Ruzzene M, Pilone MS, Pollegioni L, Molla G,
JournalJ Biol Chem
PubMed ID18544534
Human genes coding for pLG72 and d-amino acid oxidase have recently been linked to the onset of schizophrenia. pLG72 was proposed as an activator of the human FAD-containing flavoprotein d-amino acid oxidase (hDAAO). In the brain this oxidizes d-serine, a potent activator of N-methyl-d-aspartate receptor. We have investigated the mechanistic ... More
An enhanced butyrylcholinesterase method to measure organophosphorus nerve agent exposure in humans.
AuthorsPantazides BG, Watson CM, Carter MD, Crow BS, Perez JW, Blake TA, Thomas JD, Johnson RC
Journal
PubMed ID24604326
Organophosphorus nerve agent (OPNA) adducts to butyrylcholinesterase (BChE) can be used to confirm exposure in humans. A highly accurate method to detect G- and V-series OPNA adducts to BChE in 75 µL of filtered blood, serum, or plasma has been developed using immunomagnetic separation (IMS) coupled with liquid chromatography tandem mass ... More
High-affinity recombinant antibody fragments (Fabs) can be applied in peptide enrichment immuno-MRM assays.
AuthorsWhiteaker JR, Zhao L, Frisch C, Ylera F, Harth S, Knappik A, Paulovich AG
Journal
PubMed ID24568200
High-affinity antibodies binding to linear peptides in solution are a prerequisite for performing immuno-MRM, an emerging technology for protein quantitation with high precision and specificity using peptide immunoaffinity enrichment coupled to stable isotope dilution and targeted mass spectrometry. Recombinant antibodies can be generated from appropriate libraries in high-throughput in an ... More
Enhanced stability of blood matrices using a dried sample spot assay to measure human butyrylcholinesterase activity and nerve agent adducts.
AuthorsPerez JW, Pantazides BG, Watson CM, Thomas JD, Blake TA, Johnson RC
Journal
PubMed ID25955132
Dried matrix spots are safer to handle and easier to store than wet blood products, but factors such as intraspot variability and unknown sample volumes have limited their appeal as a sampling format for quantitative analyses. In this work, we introduce a dried spot activity assay for quantifying butyrylcholinesterase (BChE) ... More
Performance of a novel high throughput method for the determination of VX in drinking water samples.
AuthorsKnaack JS, Zhou Y, Magnuson M, Silvestri E, Johnson RC
JournalAnal Chem
PubMed ID23402290
VX (O-ethyl-S-(2-diisopropylaminoethyl) methylphosphonothioate) is a highly toxic organophosphorus nerve agent, and even low levels of contamination in water can be harmful. Measurement of low concentrations of VX in aqueous matrixes is possible using an immunomagnetic scavenging technique and detection using liquid chromatography/tandem-mass spectrometry. Performance of the method was characterized in ... More
Anti-idiotypic monobodies derived from a fibronectin scaffold.
AuthorsSullivan MA, Brooks LR, Weidenborner P, Domm W, Mattiacio J, Xu Q, Tiberio M, Wentworth T, Kobie J, Bryk P, Zheng B, Murphy M, Sanz I, Dewhurst S
JournalBiochemistry
PubMed ID23394681
Mimetics of conformational protein epitopes have broad applications but have been difficult to identify using conventional peptide phage display. The 10th type III domain of human fibronectin (FNfn10) has two extended, randomizable surface-exposed loops and might be more amenable to the identification of such mimetics. We therefore selected a library ... More
Multiplexed, Quantitative Workflow for Sensitive Biomarker Discovery in Plasma Yields Novel Candidates for Early Myocardial Injury.
AuthorsKeshishian H, Burgess MW, Gillette MA, Mertins P, Clauser KR, Mani DR, Kuhn EW, Farrell LA, Gerszten RE, Carr SA
Journal
PubMed ID25724909
We have developed a novel plasma protein analysis platform with optimized sample preparation, chromatography, and MS analysis protocols. The workflow, which utilizes chemical isobaric mass tag labeling for relative quantification of plasma proteins, achieves far greater depth of proteome detection and quantification while simultaneously having increased sample throughput than prior ... More
Anti-peptide monoclonal antibodies generated for immuno-multiple reaction monitoring-mass spectrometry assays have a high probability of supporting Western blot and ELISA.
AuthorsSchoenherr RM, Saul RG, Whiteaker JR, Yan P, Whiteley GR, Paulovich AG
Journal
PubMed ID25512614
Immunoaffinity enrichment of peptides coupled to targeted, multiple reaction monitoring-mass spectrometry (immuno-MRM) has recently been developed for quantitative analysis of peptide and protein expression. As part of this technology, antibodies are generated to short, linear, tryptic peptides that are well-suited for detection by mass spectrometry. Despite its favorable analytical performance, ... More
Quantitation of ortho-cresyl phosphate adducts to butyrylcholinesterase in human serum by immunomagnetic-UHPLC-MS/MS.
AuthorsJohnson D, Carter MD, Crow BS, Isenberg SL, Graham LA, Erol HA, Watson CM, Pantazides BG, van der Schans MJ, Langenberg JP, Noort D, Blake TA, Thomas JD, Johnson RC
Journal
PubMed ID26149113
Tri-ortho-cresyl phosphate (ToCP) is an anti-wear, flame retardant additive used in industrial lubricants, hydraulic fluids and gasoline. The neurotoxic effects of ToCP arise from the liver-activated metabolite 2-(o-cresyl)-4H-1,3,2-benzodioxaphosphoran-2-one (cresyl saligenin phosphate or CBDP), which inhibits esterase enzymes including butyrylcholinesterase (BChE). Following BChE adduction, CBDP undergoes hydrolysis to form the aged ... More
Direct quantitation of methyl phosphonate adducts to human serum butyrylcholinesterase by immunomagnetic-UHPLC-MS/MS.
AuthorsCarter MD, Crow BS, Pantazides BG, Watson CM, Thomas JD, Blake TA, Johnson RC
Journal
PubMed ID24205842
Hydrolysis of G- and V-series organophosphorus nerve agents (OPNAs) containing a phosphorus-methyl bond yields a methylphosphonic acid (MeP) product when adducted to human butyrylcholinesterase (BChE). The MeP adduct is considered a sign of "aging" and results in loss of the o-alkyl identifier specific to each nerve agent. After aging has ... More
Interlaboratory evaluation of automated, multiplexed peptide immunoaffinity enrichment coupled to multiple reaction monitoring mass spectrometry for quantifying proteins in plasma.
AuthorsKuhn E, Whiteaker JR, Mani DR, Jackson AM, Zhao L, Pope ME, Smith D, Rivera KD, Anderson NL, Skates SJ, Pearson TW, Paulovich AG, Carr SA
JournalMol Cell Proteomics
PubMed ID22199228
The inability to quantify large numbers of proteins in tissues and biofluids with high precision, sensitivity, and throughput is a major bottleneck in biomarker studies. We previously demonstrated that coupling immunoaffinity enrichment using anti-peptide antibodies (SISCAPA) to multiple reaction monitoring mass spectrometry (MRM-MS) produces Immunoprecipitation MRM-MS (immuno-MRM-MS) assays that can ... More
Novel dual-mode immunomagnetic method for studying reactivation of nerve agent-inhibited butyrylcholinesterase.
AuthorsAbney CW, Knaack JL, Ali AA, Johnson RC
JournalChem Res Toxicol
PubMed ID23656164
A novel immunomagnetic method has been developed for the simultaneous measurement of organophosphorus nerve agent (OPNA) adducts to butyrylcholinesterase (BuChE) and free OPNAs in serum. This new approach, deemed dual-mode immunomagnetic analysis (Dual-Mode IMA), combines immunomagnetic separation (IMS) and immunomagnetic scavenging (IMSc) and has been used to measure the effectiveness ... More
Targeted MS Assay Predicting Tamoxifen Resistance in Estrogen-Receptor-Positive Breast Cancer Tissues and Sera.
AuthorsDe Marchi T, Kuhn E, Dekker LJ, Stingl C, Braakman RB, Opdam M, Linn SC, Sweep FC, Span PN, Luider TM, Foekens JA, Martens JW, Carr SA, Umar A
JournalJ Proteome Res
PubMed ID26958999
'We recently reported on the development of a 4-protein-based classifier (PDCD4, CGN, G3BP2, and OCIAD1) capable of predicting outcome to tamoxifen treatment in recurrent, estrogen-receptor-positive breast cancer based on high-resolution MS data. A precise and high-throughput assay to measure these proteins in a multiplexed, targeted fashion would be favorable to ... More
Cerebrospinal Fluid Amyloid Beta and Tau Concentrations Are Not Modulated by 16 Weeks of Moderate- to High-Intensity Physical Exercise in Patients with Alzheimer Disease.
Physical exercise may have some effect on cognition in patients with Alzheimer disease (AD). However, the underlying biochemical effects are unclear. Animal studies have shown that amyloid beta (Aß), one of the pathological hallmarks of AD, can be altered with high levels of physical activity. ... More
A high-throughput UHPLC-MS/MS method for the quantification of five aged butyrylcholinesterase biomarkers from human exposure to organophosphorus nerve agents.
AuthorsGraham LA, Johnson D, Carter MD, Stout EG, Erol HA, Isenberg SL, Mathews TP, Thomas JD, Johnson RC
JournalBiomed Chromatogr
PubMed ID27572107
Organophosphorus nerve agents (OPNAs) are toxic compounds that are classified as prohibited Schedule 1 chemical weapons. In the body, OPNAs bind to butyrylcholinesterase (BChE) to form nerve agent adducts (OPNA-BChE). OPNA-BChE adducts can provide a reliable, long-term protein biomarker for assessing human exposure. A major challenge facing OPNA-BChE detection is ... More
A U1 snRNP-specific assembly pathway reveals the SMN complex as a versatile hub for RNP exchange.
AuthorsSo BR, Wan L, Zhang Z, Li P, Babiash E, Duan J, Younis I, Dreyfuss G
JournalNat Struct Mol Biol
PubMed ID26828962
Despite equal snRNP stoichiometry in spliceosomes, U1 snRNP (U1) is typically the most abundant vertebrate snRNP. Mechanisms regulating U1 overabundance and snRNP repertoire are unknown. In Sm-core assembly, a key snRNP-biogenesis step mediated by the SMN complex, the snRNA-specific RNA-binding protein (RBP) Gemin5 delivers pre-snRNAs, which join SMN-Gemin2-recruited Sm proteins. ... More