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991.
Summary Three techniques for measuring plasmid stability in yeasts are described and evaluated. The yeast used was aKluyveromyces lactis strain which was transformed with a plasmid, pCR1, to enable production of heterologous α-amylase. The techniques were based
on plate counts on a selective antibiotic-containing medium and a non-selective medium, and on clearing zones on starch-supplemented
plates for α-amylase detection. The plate ratio and clearing zones methods gave comparable results while the transfer colony
method estimated much lower plasmid stabilities. 相似文献
992.
993.
Graham J. Moore Renee C. Ganter John M. Matsoukas John Hondrelis George Agelis Klemoenis Barlos Scott Wilkinson John Sandall Patrick Fowler 《Journal of molecular recognition : JMR》1994,7(4):251-256
A triad of interacting group (TyrOH? His$ \underline\ominus$O2C) in angiotensin II (ANG II) has been postulated to create the tyrosinate anion pharmacophore (tyanophore) responsible for receptor activation/triggering (Biochim. Biophys. Acta 1991, 1065, 21). In the present study we investigated the effects on bioactivity of substituting the Tyr4 residue in [Sar1]ANG II with other anionic or electronegative amino acids, and with a number of aromatic amino acids lacking a hydroxyl group. [Sar1 Nva(δ-OH)4]ANG II, [Sar1 Nva(δ-OCH3)4]ANG II, [Sar1 Met4]ANG II, [Sar1 Gln4]ANG II, [Sar1 Glu4]ANG II and [Sar1 DL -Alg4]ANG II had agonist activities in the rat isolated uterus assay of 4, 3, 19, 10, > 0.1 and > 0.1%, respectively, of that of ANG II. [Sar1 Nal4]ANG II, [Sar1 Pal4]ANG II, [Sar1 DL -Phg(4′-F)4]ANG II, [Sar1 Phe(4′-F)4]ANG II, [Sar1 Phe(F5)4]ANG II and [Sar1 His4]ANG II had agonist activities of 4.5, 7, < 0.1, 0.2, 1 and 0.6%, respectively. All peptides investigated were devoid of measurable antagonist activity except [Sar1] Phe(4′-F)4 ANG II (pA2 = 7.7). These findings illustrate that anionic or electronegative aliphatic side chains replacing tyrosinate at position 4 can partially activate the angiotension receptor. For ANG II analogues containing an aromatic amino acid other than Tyr at position 4, ligand binding and agonist activity are not dependent on the electronegativity or dipole moment of the aromatic ring, or on the ability of the 4′ ring substituent to accept a proton. Modelling based on ab initio calculations of aromatic ring multipoles illustrate that the apparent binding affinity (PA2) of ANG II analogues is associated with a perpendicular electrostatic interaction of the position 4 aromatic ring with a receptor-based group. In addition, intramolecular interactions providing for the conformation of the ligand as it approaches its receptor appear to have a role in determining agonist vs antagonist activity. 相似文献
994.
Frances Jurnak Marilyn D. Yoder Richard Pickergill John Jenkins 《Current opinion in structural biology》1994,4(6)
A new type of structural domain, composed of parallel β-strands folded into a coiled structure, has been observed in several protein structures within the past year. An analysis of the basic motif indicates that there are two distinct types, with variations likely to be discovered in the future. 相似文献
995.
996.
Warden J 《BMJ (Clinical research ed.)》1994,308(6936):1062
997.
John A. Lowe III Weimin Qian Pamela J. Scott Stafford McLean Dianne K. Bryce Rosemary T. Crawford Jon Bordner 《Bioorganic & medicinal chemistry letters》1994,4(24):2877-2882
A series of 5,7-diphenyl-3-ureidohexahydroazepin-2-one cholecystokinin-B (CCK-B) receptor antagonists was synthesized using Beckmann ring expansion of a suitable 2,4-diphenylcyclohexanone as a key step. SAR studies revealed the importance of the 5-aryl group for high and selective CCK-B receptor affinity, as illustrated in compound (−)-10i (CCK-B IC50 = 6.8 nM). 相似文献
998.
We have investigated targeting to the endoplasmic reticulum (ER) of wild-type GUS and a modified form (GUS S358) by making an N-terminal fusion of the -glucuronidase (GUS) enzyme with the wheat -amylase signal peptide.In vitro studies demonstrated that the modified GUS (S358) lacked the glycosylation site present within the wild-type enzyme. Analysis of transgenic tobacco plants revealed that the modified GUS enzyme retained activity upon passage to the ER. When further experiments were carried out to determine the cellular location of the modified GUS enzyme, it was found that (contrary to expectation) the majority of GUS activity was retained within the cell and was not secreted to the cell surface via the default pathway. The data indicated that the modified GUS enzyme is an unsuitable reporter enzyme for studying protein secretion. 相似文献
999.
5e-tert-Butyl-2e-[4-(substituted-ethynyl)phenyl]-1,3-dithianes with selected functional groups (R) on the ethynyl moiety are potent blockers of the GABA-gated chloride channel measured as inhibitor concentration (IC50) for 4-n-[3H]propyl-1-(4-ethynylphenyl)-2, 6,7-trioxabicyclo[2.2.2]octanebinding to bovine brain membranes. The terminal R substituents were introduced by coupling 5e-tert-butyl-2e-(4-iodophenyl)-1,3-dithiane with HC ≡ CR or 5e-tert-butyl-2e-(4-ethynylphenyl)-1,3-dithiane with XR. The potency of the parent compound (R=H) with an IC50 of 21 μM is equaled or exceeded by up to 7-fold (i.e. IC50 = 3–21 μM) by several carboxylic acids [R = (CH2)nCO2H (n = 0–3), (CH2nOCH2CO2H (n = 1–3) and CH2SCH2 CO2H] and their esters and two phosphonic acids (CH2CH2PO3H2 and CH2OCH2PO3H2) but not their esters. These carboxyl and phosphonic acids (and their salts) include the most potent water-soluble chloride channel blockers known. Conversion to the monosulfones increases activity of the R = H and CH2OH analogs by 1.2- to 3-fold but decreases that of the R = CH2CH2CO2R′ (R′ = H or CH3) derivatives by 3- to 13-fold. Quantitative structure-activity analyses for 44 2-[4-(substituted-ethynyl)phenyl]-dithianes suggests that the principal feature of the R substituent for high activity is its polarizable volume modeled as molecular refractivity, i.e. this substituent is not a well-defined pharmacophore and undergoes a structurally non-specific interaction with the receptor. These observations lay the background for preparing candidate affinity probes. 相似文献
1000.
Lieve Vits Kristel De Boulle Edwin Reyniers Ingrid Handig John K. Darby Ben Oostra Patrick J. Willems 《Human genetics》1994,94(5):523-526
The fragile X syndrome is the result of amplification of a CGG trinucleotide repeat in the FMR1 gene and anticipation in this disease is caused by an intergenerational expansion of this repeat. Although regression of a CGG repeat in the premutation range is not uncommon, regression from a full premutation (>200 repeats) or premutation range (50–200 repeats) to a repeat of normal size (<50 repeats) has not yet been documented. We present here a family in which the number of repeats apparently regressed from approximately 110 in the mother to 44 in her daughter. Although the CGG repeat of the daughter is in the normal range, she is a carrier of the fragile X mutation based upon the segregation pattern of Xq27 markers flanking FMR1. It is unclear, however, whether this allele of 44 repeats will be stably transmitted, as the daughter has as yet no progeny. Nevertheless, the size range between normal alleles and premutation alleles overlap, a factor that complicates genetic counseling. 相似文献