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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Optimal activation of T cells to clonally expand requires at least two distinct biological signals; one is generated by the interaction of the T cell receptor (TcR) with peptides bound to MHC molecules. The other signal(s) is (are) generated by a functionally defined event called the co-stimulatory pathway. We have characterized the co-stimulatory property of a murine B lymphocyte membrane protein (155-160 kD) on resting CD4+ T cells. The study involved the isolation of a 155-160 kD protein (B1) from the membranes of LPS-stimulated B cells. When reconstituted into lipid vesicles, B1 exerted a dose-dependent proliferative response to CD4+ T cells, resulting in the predominant secretion of IL-4 and IL-5 after cross-linking receptors with anti-CD3 mAb. This protein is a phosphoglycoprotein which gives a single spot on two-dimensional gel electrophoresis under reducing conditions and as a distinct peak on reverse phase-HPLC. The B1 binds to the T cell surface as is demonstrated by electron microscopic autoradiography and scanning electron microscopy, as well as competitive binding assays. It does not cross-react with antibodies directed against ICAM-1, LFA-1 alpha, B7, HSA and VCAM-1, suggesting the novelty of the protein. Activation of CD4+ T cells with B1 in the presence of anti-CD3 resulted in the translocation of protein kinase C (PKC). The B1 is barely detectable on the surface of resting B cells and digestion of this protein with V8 protease and peptide N-glycosidase F resulted in distinct protein bands on an autoradiogram.
Mol Immunol 1996 Jan
PMID:Characterization of a novel co-stimulatory molecule: a 155-160kD B cell surface protein provides accessory help to CD4+ T cells to proliferate and differentiate. 860 18

The LH/CG receptor (LH/CG-R) is a G protein-coupled receptor with a relatively large glycosylated extracellular domain. The complete 674 amino acid residue rat receptor was expressed in Sf9 insect cells using the baculovirus expression system. Optimal expression under the control of the polyhedrin promoter was obtained at 72 h post-infection and a multiplicity of infection of 0.1. The recombinant LH/CG-R was expressed on the cell surface (ca. 4500 receptors/cell) and exhibited saturable, high affinity binding of human CG (hCG) with a Kd of 0.4 nM. There was no evidence of intracellular trapping of the receptor. The intracellular concentration of cAMP was increased in response to hCG binding. In contrast, baculovirus-expressed recombinant hCG only weakly stimulated intracellular cAMP levels at relatively high doses. Two forms of the receptor (approximately 75 and approximately 200 kDa) were detected by Western blot analysis. These results demonstrate that the full length LH/CG-R expressed in insect cells is functional in that it binds hormone with high affinity and is able to couple to adenylate cylase.
Mol Cell Endocrinol 1996 Mar 01
PMID:Expression of functional lutropin/choriogonadotropin receptor in the baculovirus system. 873 77

We utilized reverse transcription-polymerase chain reaction (RT-PCR) to examine the myocardial mRNA expression of growth factors in mice and children. Total cellular RNA was extracted from these tissues using acidified-phenol guanidinium thiocyanate. Optimal oligonucleotide primer pairs for RT-PCR were selected with the aid of the computer program Oligo 4.0. RT-PCR analysis of total cellular RNA demonstrated the measurable presence of the mRNA for the following growth factors in the myocardium of both mice and children: acidic fibroblast growth factor (aFGF), basic FGF (bFGF), insulin-like growth factor (IGF)-1, IGF-2, platelet-derived growth factor (PDGF)-A chain, PDGF-B chain, transforming growth factor (TGF)beta-1, TGFbeta-2, and TGFbeta-3. The amplified cDNA message for the growth factors and beta-actin migrated as a discrete band at the expected base pair number on agarose gels stained with the intercalating fluorescent dye ethidium bromide. Densitometry of the photographic negatives of these gels permitted the rapid and semiquantitative comparison of these factors. These data demonstrate the feasibility and reproducibility of utilizing RT-PCR for the specific detection and semiquantitation of mRNA expression of myocardial aFGF, bFGF, PDGF-A chain, PDGF-B chain, lGF-1, lGF-2, TGFbeta-1, TGFbeta-2, and TGFbeta-3 in mice and children.
Biochem Mol Med 1996 Feb
PMID:Murine and pediatric myocardial growth factor mRNA expression using reverse transcription-polymerase chain reaction. 881 19

Threading algorithms attempt to solve the inverse protein folding problem: given a group of structures and a sequence, identify the structure that is most compatible with this sequence. A recent study of this class of algorithms by S. J. Wodak and colleagues suggests that while threading algorithms are capable of recognizing many folding motifs, their performance in truly blind predictions is disappointing, and the underlying alignments upon which the selections are based are frequently errant. To help overcome this problem we have developed a Test of Optimal Mutagenesis algorithm (TOM) that exploits information inherent in the variation between several homologues in a multiple sequence alignment. This information is used to help select the correct structural motif for the sequence from a database of known structures. A total of 305 high-resolution structures were selected to represent the set of known folds; 56 proteins were chosen that had at least one close structural match in this set. To test TOM, we attempted to determine which of the 305 folds was a match to each of the 56 protein sequences. TOM correctly predicts a close structural match for 45% of these proteins. THREADER, an algorithm chosen as a literature standard, correctly matched 20% of the test set. By comparing the performance of TOM, THREADER, and TOM NOVAR (a version of TOM without variability information), we conclude that the tendency of an amino acid to be buried or exposed is the dominant determinant of the success of threading algorithms. In addition, the structural alignments produced by TOM suggest that the exact alignment of just 30 to 50% of the residues in a sequence with the correct fold is necessary to select it as the highest scoring match in a set of folds.
J Mol Biol 1996 Sep 20
PMID:Multiple sequence information for threading algorithms. 883 96

It has been shown that the novel gel matrix, PCR Purity Plus which consists of a vinyl-polymer of polyacrylamide, provides a superior and rapid means of separating DNA fragments. As this product has been discontinued, the two new commercial versions of this matrix (MDE and GeneAmp) with PCR Purity Plus were compared. Optimal conditions for resolving DNA fingerprint profiles for both matrices were defined. Both MDE and GeneAmp gels provided a clear separation of DNA fragments. However, the profiles obtained on GeneAmp gel were closest to that of PCR Purity Plus. These results should be useful to DNA fingerprinting studies where it is critical to obtain a clear resolution of complex DNA profiles.
Mol Biotechnol 1996 Feb
PMID:An evaluation of new gel matrices for the separation of PCR-amplified DNA fragments. 885 17

Functional 3 beta-hydroxysteroid dehydrogenase coupled with isomerase (3 beta-HSD) was extracted from dog pancreatic mitochondria by treatment with the zwitterionic detergent CHAPSO. Increasing concentrations of this detergent led to a progressive and simultaneous solubilization of the pregnene (C-21) and androstene (C-19) dehydrogenase activities. Optimal solubilization of both C-21 and C-19 3 beta-HSD activities was achieved at a detergent/protein ratio of 0.6 (w/w). One hundred thirty percent of the initial particulate enzyme activities were recovered in the 105,000 g supernatant fluid with a 2.5-fold increase in the enzymatic specific activities. The C-21/C-19 activity ratios were 1.3 for mitochondria and 1.39 for the solubilized preparation. The apparent Km values for steroid substrates were unchanged after solubilization. Treatment of the mitochondrial suspension with sodium deoxycholate, CTAB, Lubrol XW, Brij 58, Emulgen 913 and Triton X-100 markedly decreased the 3 beta-HSD activities as a function of the detergent concentration and failed in to achieve solubilization.
Comp Biochem Physiol B Biochem Mol Biol 1996 Oct
PMID:Detergent solubilization of 3 beta-hydroxysteroid dehydrogenase from dog pancreas. 893 6

Optimal activation of T cells requires at least two signals delivered by the T-cell receptor complex and costimulatory molecules such as CD28. The CD28 signaling participates in the transcription of the interleukin-2 gene through activation of an enhancer termed the CD28-responsive element (CD28RE). Stimulation of CD28 enhances mitogen-mediated induction of CD28RE-binding proteins including members of the NF-kappaB/Rel transcription factor family, although the underlying mechanism remains elusive. In this report, we show that CD28 costimulation leads to biphasic induction of NF-kappaB/Rel heterodimers, including early-phase induction of p50/RelA and c-Rel/RelA and late-phase induction of p50/c-Rel. Interestingly, activation of these NF-kappaB/Rel complexes by the CD28 signal is associated with the rapid degradation of both IkappaBalpha and IkappaBbeta, two major cytoplasmic inhibitors of NF-kappaB/Rel. Although IkappaBalpha degradation can be induced by phorbol ester alone, degradation of IkappaBbeta is largely dependent on the CD28 costimulatory signal. We further demonstrate that CD28-mediated transactivation of the CD28RE enhancer is potently inhibited by an N-terminal truncation mutant of IkappaBbeta that is incapable of responding to the degradation signals. Together, these results suggest that the CD28 costimulatory signal augments activation of NF-kappaB/Rel by promoting degradation of IkappaBbeta as well as enhancing degradation of IkappaBalpha and that induction of NF-kappaB/Rel serves as an essential step in the signal-mediated activation of the CD28RE enhancer.
Mol Cell Biol 1996 Dec
PMID:CD28 mediates a potent costimulatory signal for rapid degradation of IkappaBbeta which is associated with accelerated activation of various NF-kappaB/Rel heterodimers. 894 28

Using a sensitive homologous pairing/DNA strand transfer assay, we detected formation of joint molecules in the presence of nuclear extract from cultured mosquito C7-10 cells in a reaction containing single stranded circular m13 DNA and a linear, double stranded DNA 5'-end-labeled on the strand complementary to a portion of the single-stranded substrate. Joint molecules were detected by the reduced electrophoretic mobility of labeled probe on agarose gels, which indicated that the 5'-end labeled strand of the linear duplex had formed a hybrid with the single-stranded substrate. Characterization of the activity detected initially in crude nuclear extracts provided a basis for a 5-fold enrichment of activity after a two-step KCl elution from heparin-Sepharose. Further purification by preparative electrophoresis yielded a band at approximately 35 kDa, which, when transferred to Immobilon P membrane, specifically bound the labeled, complementary strand probe. Optimal activity of the electroeluted enzyme required both magnesium and ATP and was sensitive to the ratio of single-stranded and double-stranded DNA substrate and to the amount of protein. This homologous pairing activity from mosquito cells is the first such activity to be described from an insect other than Drosophila melanogaster.
Insect Biochem Mol Biol 1996 Jul
PMID:Evidence for a DNA homologous pairing activity in nuclear extracts from mosquito cells. 899 89

The possibility of using polymerase chain reaction (PCR) for the laboratory diagnosis of brucellosis was tested with a representative regional collection of 44 Brucella strains. The strains differed by the test and source of isolation, virulence, and phenotypical characteristics. Optimal parameters of PCR have been defined and the possibility of detecting the agents in pathological material from cattle demonstrated.
Mol Gen Mikrobiol Virusol
PMID:[Optimization of Brucella detection using polymerase chain reaction]. 899 21

Trophozoites of Entamoeba invadens were able to ingest glucopolysaccharides and metabolize them. An activity capable of degrading these substrates was purified to apparent homogeneity. The enzyme was identified as beta-amylase (alpha-1,4-D-glucan maltohydrolase EC 3.2.1.2): It was active against glycogen, amylose and amylopectin in a ratio of 100:198:133 and was also able to attack linear alpha-1,4-glucooligosaccharides with more than three glucose moieties. All degradation experiments yielded maltose as reaction product, and no free glucose could be detected. While amylose was completely degraded, amylolysis of glycogen and amylopectin yielded limit dextrins besides maltose. The enzyme was completely inactive against cyclohexaamylose, cycloheptaamylose and pullulan, indicating its lack of endo-glucosidase specificity. Hydrolysis of 4-nitrophenyl-maltoheptaoside resulted in the successive removal of maltose units from the non-reducing end yielding 4-nitrophenyl-maltopentaoside, -trioside and -glucoside. No 4-nitrophenyl-glycosides with even numbered glucose moieties were formed from this substrate. The enzyme exhibited a relative molecular mass of M(r) = 45,000 +/- 5% by gel filtration and sodium dodecyl sulfate (SDS) gel electrophoresis and the N-terminal sequence 1 VEVNVMLPL 9. Optimal hydrolysis was observed at pH 5.5 and a temperature of 42 degrees C. On the basis of inhibition by mercury ions and p-chloro-mercurybenzoate and abrogation of this effect by thiol reagents beta-amylase was identified as sulfhydryl-enzyme.
Mol Biochem Parasitol 1996 Dec 20
PMID:Detection, purification and partial characterization of beta-amylase from trophozoites of Entamoeba invadens. 902 50


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