Peptide separations by slab gel electrophoresis in pluronic F127 polymer liquid crystals.
AuthorsRill RL, Al-Sayah MA
JournalElectrophoresis
PubMed ID15174045
'Proteomics and peptidomics could benefit from simple methods for high-resolution separation of oligopeptides analogous to slab gel electrophoresis of proteins. Gels of Pluronic F127 copolymer surfactant were investigated as media for slab gel electrophoresis of oligopeptides using a trypsin digest of myoglobin. Concentrated solutions of Pluronic F127 are fluid at ... More
Fluorophores at the N terminus of nascent chloramphenicol acetyltransferase peptides affect translation and movement through the ribosome.
AuthorsRamachandiran V, Willms C, Kramer G, Hardesty B
JournalJ Biol Chem
PubMed ID10636875
'Structurally different fluorescent probes were covalently attached to methionyl-tRNA(f) and tested for their incorporation into nascent peptides and full-length protein using an Escherichia coli cell-free coupled transcription/translation system. Bovine rhodanese and bacterial chloramphenicol acetyltransferase (CAT) were synthesized using derivatives of cascade yellow, eosin, pyrene, or coumarin attached to [(35)S]Met-tRNA(f). All ... More
Resolution of a signal transfer region from a general binding domain in gbeta for stimulation of phospholipase C-beta2.
AuthorsBuck E, Li J, Chen Y, Weng G, Scarlata S, Iyengar R
JournalScience
PubMed ID10037604
'Signaling by guanine nucleotide-binding proteins (G proteins) involves sequential protein-protein interactions. G protein-betagamma subunit (Gbetagamma) interactions with phospholipase C-beta2 (PLC-beta2) were studied to determine if all Gbeta contacts are required for signaling. A peptide encoding Gbeta amino acid residues 86 to 105 stimulated PLC-beta2. Six residues (96 to 101) within ... More
Initiation of protein synthesis with fluorophore-Met-tRNA(f) and the involvement of IF-2.
AuthorsMcIntosh B, Ramachandiran V, Kramer G, Hardesty B
JournalBiochimie
PubMed ID10727773
'The complicity of initiation factor 2 (IF-2) in causing the observed low incorporation of N-terminal fluorophore from fluorophore-methionyl-tRNA(f) during protein synthesis in an in vitro coupled transcription/translation system was investigated. The low incorporation in comparison to formyl-methionine was not due to the lack of interaction of fluorophore-Met-tRNA(f) with IF-2. Fluorescence ... More
CD4+ T cells derived from B cell-deficient mice inhibit the establishment of peripheral B cell pools.
AuthorsBaumgarth N, Jager GC, Herman OC, Herzenberg LA
JournalProc Natl Acad Sci U S A
PubMed ID10781082
'We demonstrate that adoptive transfer of peritoneal cavity B cells fails to replenish the peripheral B-1 cells in adult B cell-deficient (mu(-/-)) mice but does replenish adult RAG-1(-/-) mice. We show that this lack of self-replenishment in mu(-/-) mice is mediated by strongly inhibitory, radiation-sensitive CD4(+) T cells that also ... More
11-color, 13-parameter flow cytometry: identification of human naive T cells by phenotype, function, and T-cell receptor diversity.
AuthorsDe Rosa SC, Herzenberg LA, Herzenberg LA, Roederer M
JournalNat Med
PubMed ID11175858
Peptide-based fluorescent sensors of protein kinase activity: design and applications.
AuthorsSharma V, Wang Q, Lawrence DS,
JournalBiochim Biophys Acta
PubMed ID17881302
Protein kinases control the flow of information through cell-signaling pathways. A detailed analysis of their behavior enhances our ability to understand normal cellular states and to devise therapeutic interventions for diseases. The design and application of ... More
"Rainbow" reporters for multispectral marking and lineage analysis of hematopoietic stem cells.
AuthorsHawley TS, Telford WG, Hawley RG
JournalStem Cells
PubMed ID11239166
Hematologic diseases potentially benefiting from gene-based therapies involving hematopoietic stem cells (HSCs) include hereditary hemoglobinopathies, immunodeficiency syndromes, and congenital bleeding disorders such as hemophilia A, as well as acquired diseases like AIDS. Successful treatment of these blood diseases with gene-modified HSCs requires high efficiency gene delivery to the target cell ... More
Pairs of violet-light-excited fluorochromes for flow cytometric analysis.
AuthorsAnderson MT, Baumgarth N, Haugland RP, Gerstein RM, Tjioe T, Herzenberg LA, Herzenberg LA
JournalCytometry
PubMed ID9845438
We describe pairs of fluorochromes for use with the 407-nm line of a violet-light-enhanced krypton ion laser. These fluorochromes and a previously described violet-light-excited reporter variant, GFP-Vex, fall into two emission classes: blue for Cascade Blue, and green/yellow for Cascade Yellow, Lucifer Yellow, and GFP-Vex. Cascade Yellow is a new ... More
Simultaneous measurement of multiple active kinase states using polychromatic flow cytometry.
AuthorsPerez OD, Nolan GP
JournalNat Biotechnol
PubMed ID11821861
Intracellular assays of signaling systems have been limited by an inability to correlate functional subsets of cells in complex populations on the basis of active kinase states. Such correlations could be important in distinguishing changes in signaling status that arise in rare cell subsets during functional activation or in disease ... More
B-1 and B-2 cell-derived immunoglobulin M antibodies are nonredundant components of the protective response to influenza virus infection.
AuthorsBaumgarth N, Herman OC, Jager GC, Brown LE, Herzenberg LA, Chen J
JournalJ Exp Med
PubMed ID10899913
We have studied the role of secreted immunoglobulin (Ig)M in protection from infection with influenza virus and delineated the relative contributions of B-1 versus B-2 cell-derived IgM in this process. Mice deficient in secreted IgM but capable of expressing surface IgM and secreting other Ig classes show significantly reduced virus ... More
Identification of a germ-line pro-B cell subset that distinguishes the fetal/neonatal from the adult B cell development pathway.
AuthorsLu LS, Tung J, Baumgarth N, Herman O, Gleimer M, Herzenberg LA, Herzenberg LA
JournalProc Natl Acad Sci U S A
PubMed ID11867763
Studies presented here show that the expression of CD4, MHC class II (Ia,) and B220 cleanly resolves a major and a minor subset within the earliest pro-B cell population (germ-line pro-B) in adult bone marrow (BM). The major subset expresses intermediate B220 and low CD4 levels. The minor subset, which ... More
Nine color eleven parameter immunophenotyping using three laser flow cytometry.
AuthorsBigos M, Baumgarth N, Jager GC, Herman OC, Nozaki T, Stovel RT, Parks DR, Herzenberg LA
JournalCytometry
PubMed ID10331625
BACKGROUND: This study describes a three laser flow cytometer, reagents, and software used to simultaneously evaluate nine distinct fluorescent parameters on one cell sample. We compare the quality of data obtained with (1) full software compensation and (2) the use of partial spectral compensation of selected pairs of parameters in ... More