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101.
Peter Schmidt Diana Lindner Cindy Montag Sandra Berndt Annette G. Beck‐Sickinger Rainer Rudolph Daniel Huster 《Biotechnology progress》2009,25(6):1732-1739
G protein‐coupled receptors (GPCRs) are a class of membrane proteins that represent a major target for pharmacological developments. However, there is still little knowledge about GPCR structure and dynamics since high‐level expression and characterization of active GPCRs in vitro is extremely complicated. Here, we describe the recombinant expression and functional folding of the human Y2 receptor from inclusion bodies of E. coli cultures. Milligram protein quantities were produced using high density fermentation and isolated in a single step purification with a yield of over 20 mg/L culture. Extensive studies were carried out on in vitro refolding and stabilization of the isolated receptor in detergent solution. The specific binding of the ligand, the 36 residue neuropeptide Y (NPY), to the recombinant Y2 receptors in micellar form was shown by several radioligand affinity assays. In competition experiments, an IC50 value in low nanomolar range could be determined. Further, a KD value of 1.9 nM was determined from a saturation assay, where NPY was titrated to the recombinant Y2 receptors. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 相似文献
102.
103.
Background
Many high-throughput genomic experiments, such as Synthetic Genetic Array and yeast two-hybrid, use colony growth on solid media as a screen metric. These experiments routinely generate over 100,000 data points, making data analysis a time consuming and painstaking process. Here we describe ScreenMill, a new software suite that automates image analysis and simplifies data review and analysis for high-throughput biological experiments. 相似文献104.
105.
106.
We analyzed the nonlinear current-voltage relationships of the early conducting state of channelrhodopsin-2 expressed in Xenopus oocytes and human embryonic kidney cells with respect to changes of the electrochemical gradients of H+, Na+/K+, and Ca2+/Mg2+. Several models were tested for wild-type ChR2 and mutations at positions E90, E123, H134, and T159. Voltage-gating was excluded as cause for the nonlinearity. However, a general enzyme kinetic model with one predominant binding site yielded good fits throughout. The empty site with an apparent charge number of about −0.3 and strong external cation binding causes some inward rectification of the uniport function. Additional inward rectification is due to asymmetric competition from outside between the transported ion species. Significant improvement of the fits was achieved by introducing an elastic voltage-divider formed by the voltage-sensitive barriers. 相似文献
107.
Gerhard Pichler Po-Yin Cheung Corinna Binder Megan O’Reilly Bernhard Schwaberger Khalid Aziz Berndt Urlesberger Georg M. Schm?lzer 《PloS one》2014,9(12)
Objective
To describe temporal changes in systolic, diastolic, and mean blood pressure (SBP, DBP, and MBP, respectively) in term and preterm infants immediately after birth.Methods
Prospective observational two-center study. In term infants SBP, DBP, and MBP were assessed non-invasively every minute for the first 15 minutes, and in preterm infants every minute for the first 15 minutes, as well as at 20, 25, 30, 45, and 60 minutes after birth. Regression analyses were performed by gender and respiratory support in all neonates; and by mode of delivery, cord clamping time, and development of ultrasound-detected brain injury in preterm neonates.Results
Term infants (n = 54) had a mean (SD) birth weight of 3298 (442) g and gestational age of 38 (1) weeks, and preterm infants (n = 94) weighed 1340 (672) g and were 30 (3) weeks gestation. Term infants’ SBP, DBP and MBP within the first 15 minutes after birth were independent of gender or respiratory support. Linear mixed regression analysis showed that preterm infants, who were female, born vaginally, had delayed cord clamping and did not require positive pressure ventilation nor develop periventricular injury or ventriculomegaly, had significantly higher SBP, DBP, and MBP at some measurement points within the first hour after birth.Conclusions
We present novel reference ranges of BP immediately after birth in a cohort of term and preterm neonates. They may aid in optimization of cardiovascular support during early transition at all gestations. 相似文献108.
G L May L C Dunlop K Sztelma M C Berndt T C Sorrell 《Biochemical and biophysical research communications》1992,183(3):1062-1069
Proton magnetic resonance spectroscopy has been used to monitor the effect of GMP-140 on the stimulation of human peripheral blood neutrophils. Stimulation of neutrophils by lipopolysaccharide gives rise to a high resolution lipid spectrum from the intact cells. Fluid phase GMP-140, which prevents adhesion and development of inflammatory responses of neutrophils, was found to inhibit these changes in the lipid spectrum by up to 40%. Anti-GMP-140 Fab fragments reversed this effect while non-immune Fab fragments did not affect the observed inhibition by GMP-140. 相似文献
109.
In the quantitation of amino acids by precolumn derivatization with phenylisothiocyanate, the yields of N'-phenylthiocarbamyl (PTC)-aspartate and PTC-glutamate from protein hydrolysates are often suboptimal, particularly in analyses following rapid hydrolysis at 160 degrees C. In this paper we show that these losses are due to the presence of materials extracted from the glass container during hydrolysis. In the presence of these extracts, the repeated drying and neutralization steps which precede phenylthiocarbamylation result in samples not fully solubilized by the presently used derivatizing mixtures. Thus the coupling yields for the acidic residues are highly variable. A coupling buffer with the composition 35% H2O, 30% acetonitrile, 25% pyridine, and 10% triethylamine (v/v/v/v) is an efficient solvent for all amino acids in hydrolysates and permits consistent, quantitative derivatization of all amino acids, including aspartate and glutamate. 相似文献
110.
Yu-Ru Su Chongzhi Di Stephanie Bien Licai Huang Xinyuan Dong Goncalo Abecasis Sonja Berndt Stephane Bezieau Hermann Brenner Bette Caan Graham Casey Jenny Chang-Claude Stephen Chanock Sai Chen Charles Connolly Keith Curtis Jane Figueiredo Manish Gala Li Hsu 《American journal of human genetics》2018,102(5):904-919