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

Influence of surrounding media on the appearance of surface structure of Candida albicans was further investigated by rapid-freezing and freeze-fracturing techniques with a scanning electron microscope. Fibrillar structure was observed on the surface of the cells treated with water, chloroform, trichloroethane, toluene or isoamyl alcohol, but not on that of the cells treated with methanol, ethanol, isopropanol, n-butanol, acetic acid or acetone. The appearance of the fibrillar structure is proposed to be discussed in a viewpoint of the chemical interactions among the constituent molecules of a fibril, water molecules bound to the fibril molecules and the molecules of surrounding medium, especially a role of water molecules bound to the fibril molecules by hydrogen bonds.
Cell Mol Biol (Noisy-le-grand) 1993 Jun
PMID:Water molecules bound to fibrils by hydrogen bonds play an important role on the surface fibrillar structure of Candida albicans cells. 832 78

Although several methods for isolating genomic DNA from trypanosomatid protozoa exist, all are time-consuming and cumbersome. Faster, simpler and efficient protocols for preparation of DNA from these protozoa are needed to ease the screening of mutants and transfectants. We describe the use of a bacterial lysis method to isolate chromosomal DNA from a wide range of trypanosomatids. The method is based on the finding reported by He et al., who noticed that phenol/chloroform treatment of Escherichia coli cells in the presence of LiCl and Triton X-100 solubilizes plasmid DNA, while precipitating unwanted chromosomal DNA and denatured cellular proteins. In applying this lysis method to the isolation of episomal DNA from transfected trypanosomatids, we found that, unlike bacterial genomic DNA, chromosomal DNA of trypanosomatids was soluble in the phenol/chloroform/Triton/LiCl mixture. This observation prompted us to use the bacterial lysis method as a routine protocol for extraction of DNA from trypanosomatids.
Mol Biochem Parasitol 1993 Jun
PMID:Rapid isolation of DNA from trypanosomatid protozoa using a simple 'mini-prep' procedure. 834 29

We describe a novel technique for isolation of sequences that are present in one genome (tracer), but absent in another (driver). Tracer DNA, cleaved with Sau 3A and capped with a single stranded PCR adapter, is allowed to hybridize with an excess of sheared biotinylated driver; biotinylated DNA and its hybrids with the tracer are removed by phenol/chloroform extraction after incubation with streptavidin. After several rounds of subtraction the ends of self-annealed tracer molecules from the nonextractable fraction are filled-in with Tag polymerase and amplified, using the single stranded PCR adapter as a primer. The method has been applied to purification of fragments from a 2.9 kb plasmid added to E. coli DNA at equimolar quantity. Plasmid derived fragments (250-1000 bp), initially comprising 1/1400th part of tracer DNA, were purified to homogeneity after two rounds of subtraction followed by PCR.
Mol Gen Mikrobiol Virusol
PMID:A method for isolation of sequences missing in one of two related genomes. 835 Aug 79

We examine here the biochemical properties and epididymal localization of a maturation dependent ram sperm surface antigen. A monoclonal antibody, ESA152, identifies an antigen that is present on the surface of ejaculated sperm, but is absent from testicular sperm. Crosslinking of the ESA152 antigen with bivalent antibodies induces the acrosome reaction, redistributing the antigen into the anterior region of the sperm head where it associates with the fusion product of the plasma membrane and the outer acrosomal membrane. The ESA152 antigen appears as a polypeptide of 18 kDa on immunoblots of SDS-polyacrylamide gels. The ESA152 epitope includes the sialic acid termini of N-linked oligosaccharides, as shown by its sensitivity to neuraminidase and endoglycosidase F. The ESA152 antigen is a highly hydrophobic integral membrane protein that resists aqueous extraction, partitions into the detergent phase of Triton-X-114, and solubilizes in chloroform-methanol mixtures. The anchoring of ESA152 is unaffected by phosphtidylinositol specific phospholipase C. The antigen is absent from extracts of caput and corpus epididymidis but appears abruptly in the first segment of the cauda. Immunofluorescence reveals that the ESA152 epitope first appears in clusters of cells in the luminal epithelium of the proximal cauda, prior to or concurrent with its appearance on sperm.
Mol Reprod Dev 1993 Jul
PMID:Biochemical characterization and epididymal localization of the maturation-dependent ram sperm surface antigen ESA152. 835 35

Superoxide dismutase functions as a scavenger of superoxide radical protecting living organisms. This enzyme has potential use as anti-inflammatory or anti-reperfusion injury drug. Here we present a simple and efficient SOD purification method from human placental blood. Superoxide dismutase from clarified haemolysed placental blood after chloroform and ethanol treatment was purified by DEAE-Sepharose, Phenyl-Sepharose chromatographies and cross flow ultrafiltration. The purified product is 98% pure by SDS-PAGE with 71% yield and specific activity of 2.8 x 10(5) U/mg protein.
Biochem Mol Biol Int 1993 May
PMID:Purification of superoxide dismutase from placental haemolisate blood: a simple and efficient method. 835 35

The M(r) 30,000 polypeptide of the hydrophobic protein fraction of the energy-transducing NADH-ubiquinone oxidoreductase (complex I) of bovine heart mitochondria was identified as the ND2 gene product based on a comparison of amino acid analysis and partial N-terminal sequencing results with the known DNA sequence of ND2 (Anderson, S. et al. (1982) J. Mol. Biol. 156, 683-717). A simple purification procedure was devised for this ND2 gene product. The procedure, which is described, involves treatment of bovine complex I with a chloroform-methanol (2:1 [v/v]) solution. The antiserum raised against this purified bovine ND2 gene product cross-reacted with the approximately M(r) 39,000 polypeptide extracted from the Paracoccus denitrificans membranes with chloroform-methanol (2:1 [v/v]).
Biochem Mol Biol Int 1993 Jun
PMID:Isolation and characterization of the ND2 polypeptide of the bovine energy-transducing NADH-ubiquinone oxidoreductase (complex I). 836 7

The nature of the inositol phosphates present in adult rat left atria have been examined. Trichloroacetic acid extracts of [3H] inositol-labelled left atria contained the 1 (or 3)-, 2- and 4-isomers of inositol monophosphate, inositol-1,4-bisphosphate, and inositol-1,4,5-trisphosphate. Addition of noradrenaline increased all of these isomers. However, even in the presence of noradrenaline, there was no evidence for the presence of inositol-1,3,4,5-tetrakisphosphate or its dephosphorylation products including inositol-1,3,4-trisphosphate and the 1,3- and 3,4-isomers of inositol bisphosphate. Total accumulation of inositol phosphates in the presence of LiCl was 98% accounted for by dephosphorylation of inositol-1,4,5-trisphosphate and its cyclic isomer. These data indicate that, in intact left atria, metabolism of inositol-1,4,5-trisphosphate is by dephosphorylation and that significant activity of the phosphorylation pathway is not present. When extractions were performed using either acidified chloroform-methanol or perchloric acid, other [3H] inositol-labelled compounds were detected in atrial extracts. One of these chromatographed with ATP and might be mistaken for inositol-1,3,4-trisphosphate if co-chromatography with ATP was used to identify that compound. Another compound had a similar, but not identical chromatographic profile to inositol-1,3,4,5-tetrakisphosphate and is thus likely to be mistaken for that compound. In addition, perchloric acid extraction caused phosphate group migration generating extra isomers of the inositol phosphates and other unidentified [3H]-labelled compounds. Such extraction artifacts are likely especially problematic in studies of intact heart tissue because of the time required to effectively homogenize the tissue. These findings demonstrate the need for caution in interpreting data relating to estimations of inositol phosphates in intact heart tissue.
J Mol Cell Cardiol 1993 Feb
PMID:Inositol phosphates in rat atria and the importance of the extraction procedure. 838 56

The fragile X(A) or FRAXA syndrome is the most common form of familial mental retardation and is associated with a fragile site at Xq27.3. The gene responsible for the FRAXA syndrome, the FMR1 gene, has been cloned. inactivation of the FMR1 gene is associated with amplification of a trinucle-otide CGG repeat sequence and methylation of an adjacent CpG island. Previous estimates for the prevalence of the FRAXA syndrome have been based on indirect methods of chromosome analysis in institutions and community workshops for the mentally handicapped. We have analyzed the frequency of premutations of the FMR1 gene in 3002 X chromosomes of 1000 male and 1000 female consecutive newborn nonautoclaved blood spots in an anonymous, unlinked survey. The CGG repeat sizes were calculated by measuring the length of products of the PCR reaction based on the molecular size of labeled markers in a denaturing sequencing gel assay. For consistent PCR amplification a DNA microextraction was necessary, including a phenol/chloroform series. In our population, the CGG allele ranged from 9 to 106 repeats: 97% of alleles had fewer than 40 repeats. The most frequent allele was a repeat of 28. Approximately 2.3% of alleles had CGG repeats ranging from 4 to 49 and 0.37% of alleles had repeats ranging from 50 to 59. The frequency of alleles > 60 repeats in the Manitoba male population is approximately 0.13%. The use of nonautoclaved Guthrie blood spots for population screening of FRAXA premutations is not recommended. The necessity of a phenol/chloroform DNA microextraction is tedious and time consuming. The low yield of DNA (250 ng) does not allow for reanalysis by Southern of apparently homozygous females with potentially unstable CGG alleles in the 40-60 repeat range and likely underestimates premutation carrier status.
Biochem Mol Med 1995 Oct
PMID:Frequency of FMR1 premutations in a consecutive newborn population by PCR screening of Guthrie blood spots. 859 39

A simple and efficient procedure was described for the isolation of total RNA from the fission yeast Schizosaccharomyces pombe. The present study demonstrated that the quality and the quantity of S. pombe RNA were increased by substituting phenol/chloroform mixture for phenol as a deproteinizing agent in the first vortexing step and using an ice bath instead of a dry ice-ethanol bath in the freezing step. Additionally, this protocol had the advantage of extracting total RNA without any degradation of S. pombe cells. Furthermore, the high amounts and quality of RNA extracted by this modified procedure enabled us to perform some experiments such as Northern blot, S1 mapping, primer extension, and reverse transcriptase reaction-polymerase chain reaction (RT-PCR) without further RNA purification. We suggest that this procedure is very useful to analyse primary structures and steady-state levels of RNA from S. pombe.
Biochem Mol Biol Int 1995 Oct
PMID:A simple and efficient method for the isolation of total RNA from the fission yeast Schizosaccharomyces pombe. 867 17

Rhizobium leguminosarum bv. trifolii, strain NA 30 nodulates both red (Trifolium pratense) and white (T. repens) clover and produces an acidic exopolysaccharide (EPS) containing glucose, galactose, glucuronic acid, acetate and ketalpyruvate residues in a 5:1:2:1:2 molar ratio. The in vitro synthesis of this EPS as well as the characterization of five structurally related lipid linked oligosaccharides is described employing EDTA treated cells as enzyme preparation and 14C-labelled UDP-Glc, UDP-GlcA, Acetyl CoA and phosphoenol pyruvate (PEP) and doubly labelled 32P UDP-14C-Glc as precursors. The lipidic derivatives, extracted with chloroform, methanol, water (1:2:0,3) had the properties expected for prenyl-diphospho-sugars, as judged by the pattern of labelling, DEAE cellulose column chromatography, catalytic reduction and acid lability, etc. The sugar moieties of these phosphoprenyl derivatives were identified as the acetylated octasaccharide repeating unit, its mono- and di-ketalpyruvate derivatives and two trisaccharides, one of them acetylated, on the basis of specific labelling, gel filtration, paper electrophoresis and chromatography, TLC, permethylation, etc. In vitro polymer synthesis was greatly increased when electroporated cells were substituted for EDTA treated cells as enzyme system.
Cell Mol Biol (Noisy-le-grand) 1996 Jul
PMID:The in vitro biosynthesis of the exopolysaccharide produced by Rhizobium leguminosarum bv. trifolii, strain NA 30. 883 6


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