AuthorsEvans PC, Smith TS, Lai MJ, Williams MG, Burke DF, Heyninck K, Kreike MM, Beyaert R, Blundell TL, Kilshaw PJ,
JournalJ Biol Chem
PubMed ID12682062
A previous report from this laboratory described two novel proteins that have sequence similarity to A20, a negative regulator of NF-kappaB (Evans, P. C., Taylor, E. R., Coadwell, J., Heyninck, K., Beyaert, R., and Kilshaw, P. J. (2001) Biochem. J. 357, 617-623). One of these molecules, cellular zinc finger anti-NF-kappaB ... More
Mass spectrometric analysis of GAP-43/neuromodulin reveals the presence of a variety of fatty acylated species.
Authors Liang Xiquan; Lu Yun; Neubert Thomas A; Resh Marilyn D;
JournalJ Biol Chem
PubMed ID12105219
'GAP-43 (neuromodulin) is a protein kinase C substrate that is abundant in developing and regenerating neurons. Thioester-linked palmitoylation at two cysteines near the GAP-43 N terminus has been implicated in directing membrane binding. Here, we use mass spectrometry to examine the stoichiometry of palmitoylation and the molecular identity of the ... More
M phase phosphoprotein 1 is a human plus-end-directed kinesin-related protein required for cytokinesis.
AuthorsAbaza A, Soleilhac JM, Westendorf J, Piel M, Crevel I, Roux A, Pirollet F,
JournalJ Biol Chem
PubMed ID12740395
'The human M phase phosphoprotein 1 (MPP1), previously identified through a screening of a subset of proteins specifically phosphorylated at the G2/M transition (Matsumoto-Taniura, N., Pirollet, F., Monroe, R., Gerace, L., and Westendorf, J. M. (1996) Mol. Biol. Cell 7, 1455-1469), is characterized as a plus-end-directed kinesin-related protein. Recombinant MPP1 ... More
A protease assay for two-photon crosscorrelation and FRET analysis based solely on fluorescent proteins.
'GFP and the red fluorescent protein, DsRed, have been combined to design a protease assay that allows not only for fluorescence resonance energy transfer (FRET) studies but also for dual-color crosscorrelation analysis, a single-molecule-based method that selectively probes the concomitant movement of two distinct tags. The measurement principle is based ... More
Disruptor of telomeric silencing-1 is a chromatin-specific histone H3 methyltransferase.
AuthorsLacoste N, Utley RT, Hunter JM, Poirier GG, Cote J
JournalJ Biol Chem
PubMed ID12097318
'Yeast disruptor of telomeric silencing-1 (DOT1) is involved in gene silencing and in the pachytene checkpoint during meiotic cell cycle. Here we show that the Dot1 protein possesses intrinsic histone methyltransferase (HMT) activity. When compared with Rmt1, another putative yeast HMT, Dot1 shows very distinct substrate specificity. While Rmt1 methylates ... More
Despite years of investigation, the molecular mechanism responsible for regulation of the c-Abl tyrosine kinase has remained elusive. We now report inhibition of the catalytic activity of purified c-Abl in vitro, demonstrating that regulation is an intrinsic property of the molecule. We show that the interaction of the N-terminal 80 ... More
Global conformational rearrangements in integrin extracellular domains in outside-in and inside-out signaling.
How ligand binding alters integrin conformation in outside-in signaling, and how inside-out signals alter integrin affinity for ligand, have been mysterious. We address this with electron microscopy, physicochemical measurements, mutational introduction of disulfides, and ligand binding to alphaVbeta3 and alphaIIbbeta3 integrins. We show that a highly bent integrin conformation is ... More
Structure of a c-Kit Product Complex Reveals the Basis for Kinase Transactivation.
AuthorsMol CD, Lim KB, Sridhar V, Zou H, Chien EY, Sang BC, Nowakowski J, Kassel DB, Cronin CN, McRee DE,
JournalJ Biol Chem
PubMed ID12824176
The c-Kit proto-oncogene is a receptor protein-tyrosine kinase associated with several highly malignant human cancers. Upon binding its ligand, stem cell factor (SCF), c-Kit forms an active dimer that autophosphorylates itself and activates a signaling cascade that induces cell growth. Disease-causing human mutations that activate SCF-independent constitutive expression of c-Kit ... More
Cluster analysis of mass spectrometry data reveals a novel component of SAGA.
AuthorsPowell DW, Weaver CM, Jennings JL, McAfee KJ, He Y, Weil PA, Link AJ,
JournalMol Cell Biol
PubMed ID15282323
The SAGA histone acetyltransferase and TFIID complexes play key roles in eukaryotic transcription. Using hierarchical cluster analysis of mass spectrometry data to identify proteins that copurify with components of the budding yeast TFIID transcription complex, we discovered that an uncharacterized protein corresponding to the YPL047W open reading frame significantly associated ... More
Studies on the Mode of Ku Interaction with DNA.
Authors Arosio Daniele; Cui Sheng; Ortega Claudia; Chovanec Miroslav; Di Marco Stefania; Baldini Giancarlo; Falaschi Arturo; Vindigni Alessandro;
JournalJ Biol Chem
PubMed ID11796732
The Ku heterodimer plays a central role in non-homologous end-joining. The binding of recombinant Ku to DNA has been investigated by dynamic light scattering, double-filter binding, fluorescence spectroscopy, and band shift assays. The hydrodynamic radius of Ku in solution is 5.2 nm and does not change when a 25-bp double-strand ... More