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1.
Katariina M. Hutterer Robert W. Hong Jonathon Lull Xiaoyang Zhao Tian Wang Rex Pei M. Eleanor Le Oleg Borisov Rob Piper Yaoqing Diana Liu Krista Petty Izydor Apostol Gregory C. Flynn 《MABS-AUSTIN》2013,5(4):608-613
Manufacturing-induced disulfide reduction has recently been reported for monoclonal human immunoglobulin gamma (IgG) antibodies, a widely used modality in the biopharmaceutical industry. This effect has been tied to components of the intracellular thioredoxin reduction system that are released upon cell breakage. Here, we describe the effect of process parameters and intrinsic molecule properties on the extent of reduction. Material taken from cell cultures at the end of production displayed large variations in the extent of antibody reduction between different products, including no reduction, when subjected to the same reduction-promoting harvest conditions. Additionally, in a reconstituted model in which process variables could be isolated from product properties, we found that antibody reduction was dependent on the cell line (clone) and cell culture process. A bench-scale model using a thioredoxin/thioredoxin reductase regeneration system revealed that reduction susceptibility depended on not only antibody class but also light chain type; the model further demonstrates that the trend in reducibility was identical to DTT reduction sensitivity following the order IgG1λ > IgG1κ > IgG2λ > IgG2κ. Thus, both product attributes and process parameters contribute to the extent of antibody reduction during production. 相似文献
2.
Matthew R. Fisk Kent G. Apostol Amy L. Ross‐Davis Dexter O. Cahoy Anthony S. Davis 《Restoration Ecology》2019,27(3):616-625
Despite advances in restoration of degraded lands around the world, native plants are still underutilized. Selection of appropriate plant materials is a critical factor in determining plant establishment and persistence. To better inform decision‐making, we examined cold‐hardiness dynamics, flowering phenology, and survival among five geographically distinct sulfur‐flower buckwheat (Polygonaceae: Eriogonum umbellatum Torr.) populations in a common garden. LT50 (a measure of freezing injury) was determined every 6 weeks across a complete year; one population was also evaluated at the source. Cold‐hardiness dynamics were similar across populations, with annual fluctuations in mean LT50 exceeding 40°C. Rate of deacclimation (i.e. loss of cold tolerance) in spring varied across populations and was not related to the elevation from which a population came. Plants were less cold hardy in October 2014 compared to October 2013, likely reflecting a response to colder local conditions in 2013. Although the range of LT50 was similar for a single comparison of common garden versus wild‐grown plants, wild‐grown plants acclimated and deacclimated earlier than common garden‐grown plants. Plants derived from a low‐elevation population showed delayed flowering phenology, while high‐elevation populations showed earlier flowering phenology, with one high‐elevation population having the lowest survival rate in the common garden. These results suggest that while considerable plasticity in seasonal cold‐hardiness dynamics occur, population variability in deacclimation and flowering phenology have implications for selection and movement of sulfur‐flower buckwheat for ecological restoration. 相似文献
3.
Marcin I. Apostol Michael R. Sawaya Duilio Cascio David Eisenberg 《The Journal of biological chemistry》2010,285(39):29671-29675
A single nucleotide polymorphism (SNP) in codon 129 of the human prion gene, leading to a change from methionine to valine at residue 129 of prion protein (PrP), has been shown to be a determinant in the susceptibility to prion disease. However, the molecular basis of this effect remains unexplained. In the current study, we determined crystal structures of prion segments having either Met or Val at residue 129. These 6-residue segments of PrP centered on residue 129 are “steric zippers,” pairs of interacting β-sheets. Both structures of these “homozygous steric zippers” reveal direct intermolecular interactions between Met or Val in one sheet and the identical residue in the mating sheet. These two structures, plus a structure-based model of the heterozygous Met-Val steric zipper, suggest an explanation for the previously observed effects of this locus on prion disease susceptibility and progression. 相似文献
4.
Crystal structure of a major secreted protein of Mycobacterium tuberculosis-MPT63 at 1.5-A resolution 下载免费PDF全文
Goulding CW Parseghian A Sawaya MR Cascio D Apostol MI Gennaro ML Eisenberg D 《Protein science : a publication of the Protein Society》2002,11(12):2887-2893
MPT63 is a small, major secreted protein of unknown function from Mycobacterium tuberculosis that has been shown to have immunogenic properties and has been implicated in virulence. A BLAST search identified that MPT63 has homologs only in other mycobacteria, and is therefore mycobacteria specific. As MPT63 is a secreted protein, mycobacteria specific, and implicated in virulence, MPT63 is an attractive drug target against the deadliest infectious disease, tuberculosis (TB). As part of the TB Structural Genomics Consortium, the X-ray crystal structure of MPT63 was determined to 1.5-Angstrom resolution with the hope of yielding functional information about MPT63. The structure of MPT63 is an antiparallel beta-sandwich immunoglobulin-like fold, with the unusual feature of the first beta-strand of the protein forming a parallel addition to the small antiparallel beta-sheet. MPT63 has weak structural similarity to many proteins with immunoglobulin folds, in particular, Homo sapiens beta2-adaptin, bovine arrestin, and Yersinia pseudotuberculosis invasin. Although the structure of MPT63 gives no conclusive evidence to its function, structural similarity suggests that MPT63 could be involved in cell-host interactions to facilitate endocytosis/phagocytosis. 相似文献
5.
Basnakian AG Apostolov EO Yin X Abiri SO Stewart AG Singh AB Shah SV 《Experimental cell research》2006,312(20):4139-4149
The invasiveness of breast cancer cells was shown to be associated with the suppressed ability to develop apoptosis. The role of cell death DNases/endonucleases has not been previously examined in relation with the invasiveness of breast cancer cells. We have compared the activity of the endonucleases in seven human breast cancer cell lines different in the level of invasiveness and differentiation. The invasiveness of cell lines was confirmed by an in vitro Matrigel-based assay. The total endonuclease activity in the differentiated non-invasive (WDNI) cell lines was higher than that in the poorly differentiated invasive (PDI) cells. The expression of EndoG strongly correlated with the degree of estrogen receptor expression and showed an inverse correlation with vimentin and matrix metalloproteinase-13. The EndoG-positive WDNI cells were more sensitive to etoposide- or camptothecin-induced cell death than EndoG-negative PDI cells. Silencing of EndoG caused inhibited of SK-BR-3 WDNI cell death induced by etoposide. Human ductal carcinomas in situ expressed high levels of EndoG, while invasive medullar and ductal carcinomas had significantly decreased expression of EndoG. This correlated with decreased apoptosis as measured by TUNEL assay. Our findings suggest that the presence of EndoG in non-invasive breast cancer cells determines their sensitivity to apoptosis, which may be taken into consideration for developing the chemotherapeutic strategy for cancer treatment. 相似文献
6.
7.
Biodegradation of a xanthene dyes was investigated for the first time using anaerobic granular sludge. On a first screening, biomass was able to decolorize, at different extents, six azo dye solutions: acid orange 7, direct black 19, direct blue 71, mordant yellow 10, reactive red 2 and reactive red 120 and two xanthene dyes--Erythrosine B and Eosin Y. Biomass concentration, type of electron donor, induction of biomass with dye and mediation with activated carbon (AC) were variables studied for Erythrosine B (Ery) as model dye. Maximum color removal efficiency was achieved with 4.71 g VSS L?1, while the process rates were independent of the biomass concentration above 1.89 g VSS L?1. No considerable effects were observed when different substrates were used as electron donors (VFA, glucose or lactose). Addition of Ery in the incubation period of biomass led to a fivefold increase of the decolorization rate. The rate of Ery decolorization almost duplicated in the presence of commercial AC (0.1 g L?1 AC?). Using different modified AC samples (from the treatment of AC?), a threefold higher rate was obtained with the most basic one, AC(H?), as compared with non-mediated reaction. Higher rates were obtained at pH 6.0. Chemical reduction using Na?S confirmed the recalcitrant nature of this dye. The results attest that decolorization of Ery is essentially due to enzymatic and adsorption phenomena. 相似文献
8.
Deletion analysis of a multifunctional yeast tRNA ligase polypeptide. Identification of essential and dispensable functional domains. 总被引:5,自引:0,他引:5
B L Apostol S K Westaway J Abelson C L Greer 《The Journal of biological chemistry》1991,266(12):7445-7455
Splicing of tRNA precursors in extracts of Saccharomyces cerevisiae requires the action of two enzymes: a site specific endonuclease and a tRNA ligase. The tRNA ligase contains three distinct enzymatic activities: a polynucleotide kinase, a cyclic phosphodiesterase, and an RNA ligase. The polypeptide also has a high affinity pre-tRNA binding site based on its ability to form stable complexes with pre-tRNA substrates. To investigate the organization of functional enzymatic and binding elements within the polypeptide a series of defined tRNA ligase gene deletions were constructed and corresponding proteins were expressed in Escherichia coli as fusions with bacterial dihydrofolate reductase (DHFR). The DHFR/ligase derivative proteins were then efficiently purified by affinity chromatography. The complete ligase fusion protein retained enzymatic and binding activities which were unaffected by the presence of the DHFR segment. Examination of tRNA ligase deletion derivatives revealed that the amino-terminal region was required for adenylylation, while the carboxyl-terminal region was sufficient for cyclic phosphodiesterase activity. Deletions within the central region affected kinase activity. Pre-tRNA binding activity was not strictly correlated with a distinct enzymatic domain. A DHFR/ligase-derived protein lacking kinase activity efficiently joined tRNA halves. We postulate that this variant utilizes a novel RNA ligation mechanism. 相似文献
9.
Julia S. Apostolova Georgi Apostolov Ljuben M. Sirakov 《Biochimica et Biophysica Acta (BBA)/General Subjects》1976,428(2):414-419
The specific binding of insulin to the membranes from lactating mouse mamary gland was studied as a model of hormonereceptor type of binding. The basic ingredients of binding, the concentration of receptor protein and the concentration of labeled insulin were mainly studied. The characteristic changes in specific binding were followed, the adequate regression equation was drawn and the optimum conditions of binding were established for further experiments. The expediency of applying shortened orthogonal plans, regression analysis and graphic conture analysis were proved. 相似文献
10.
Bharat Thyagarajan David R Jacobs Jr George G Apostol Lewis J Smith Robert L Jensen Robert O Crapo R Graham Barr Cora E Lewis O Dale Williams 《Respiratory research》2008,9(1):31