Abstract：
:Autophagy is the process of degradation of intracellular proteins through the lysosome. Members of the tripartite motif (TRIM) proteins have shown to directly recognize autophagic cargo and also to act as a hub for the phagophore nucleation complex. The TRIM proteins are classically characterized by the presence of an amino-terminal RING domain and a B-box domain followed by a coiled coil domain. Although regarded as ubiquitin E3 ligases, this activity has been shown only for a minor set of the 79 human TRIM proteins. Additionally, the role of each domain in the E3 ligase activity is unknown. We investigated the role of the SPRY and RING domains of the human TRIM49 protein in its E3 ubiquitin ligase activity. Wild-type and mutant constructs of tagged TRIM49 were expressed in E. coli or mammalian cells, and the autoubiquitination activity of the purified protein was assessed. The purified TRIM49 showed no ubiquitin E3 ligase activity in vitro. However, cells transfected with the wild-type or mutant protein showed increased levels of lower mass polyubiquitinated proteins and both proteins copurified with polyubiquitinated proteins. Taken together, these results indicate that the TRIM49 protein plays a role in autophagic protein degradation independently of an ubiquitin E3 ligase activity.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Guimarães DS,Gomes MDdoi
10.1016/j.pep.2017.10.014subject
Has Abstractpub_date
2018-03-01 00:00:00pages
57-61eissn
1046-5928issn
1096-0279pii
S1046-5928(17)30494-1journal_volume
143pub_type
杂志文章abstract：:Disulfide bonds are normally formed after a polypeptide has been exported from the reducing environment of the cytoplasm into a more oxidizing compartment, such as the bacterial periplasm. Recently, we showed that in Escherichia coli trxB gor mutants, in which the reduction of thioredoxin and glutathione is impaired, ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2001.1520
更新日期：2001-11-01 00:00:00
abstract：:Protein insolubility often poses a significant problem during purification protocols and in enzyme assays, especially for eukaryotic proteins expressed in a recombinant bacterial system. The limited solubility of replication factor C (RFC), the clamp loader complex from Saccharomyces cerevisiae, has been previously do...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2012.03.010
更新日期：2012-06-01 00:00:00
abstract：:Plasmids pQE-60 and pQE-30 containing 6 x His-tag sequence were used for expression of fragments of HCV structural and non-structural proteins in Escherichia coli (E. coli). The following fragments were used: core (1-98 aa), NS3 (202-482 aa), and tetramer of hypervariable region 1 (HVR1) of E2 protein. The constructed...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.06.011
更新日期：2006-11-01 00:00:00
abstract：:L-Threonine dehydrogenase was purified 10,000-fold to a specific activity approximately 300 mumol.min-1.mg-1 protein from porcine liver mitochondria. Purification to apparent homogeneity was achieved by sequential chromatography on DEAE Sepharose FF, Affi-Gel Blue, Sephacryl S-200, Matrex Gel Red A, and Matrex Gel Gre...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1994.1061
更新日期：1994-10-01 00:00:00
abstract：:The guanine nucleotide exchange factor EF-1beta gene from the thermoacidophilic archaeon Sulfolobus solfataricus (SsEF-1beta) was amplified by PCR and cloned into the pT7-7 expression vector. One of four selected clones harbored the T160C nucleotide substitution leading to the Y54H amino acid change in a hydrophobic r...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1997.0806
更新日期：1998-02-01 00:00:00
abstract：:In this work we communicate the heterologous expression of a laccase from Coriolopsis gallica in Pichia pastoris. This enzyme has been reported to efficiently degrade a variety of pollutants such as industrial dyes. The expression strategy included using a previously reported modified α-factor preproleader for enhance...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2017.06.001
更新日期：2017-08-01 00:00:00
abstract：:Cell extracts of Clostridium kluyveri grown on ethanol plus succinate contained a NAD(H) dependent 4-hydroxybutanoate dehydrogenase (EC 1.1.1.61) at 66 U/mg. This enzyme was purified 42-fold under anaerobic conditions to homogeneity. Heat treatment, ion exchange chromatography on DEAE-cellulose, nondenaturing polyacry...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1995.1026
更新日期：1995-04-01 00:00:00
abstract：:G-protein coupled receptors (GPCRs) are seven transmembrane helical proteins involved in cell signaling and response. They are targets for many existing therapeutic agents, and numerous drug discovery efforts. Production of large quantities of these receptors for drug screening and structural biology remains challengi...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.10.017
更新日期：2007-04-01 00:00:00
abstract：:The transcription factor IIIC (TFIIIC) is a multisubunit DNA-binding factor required for promoter recognition and TFIIIB assembly on tRNA genes transcribed by RNA polymerase III. Yeast TFIIIC consists of six subunits, organized in the two globular subcomplexes tauA and tauB, which recognize two internal tDNA promoter ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.06.013
更新日期：2006-02-01 00:00:00
abstract：:Malassezia globosa is pathogenic fungus that causes skin disorders including dandruff in humans. Many yeast cytochrome CYP enzymes are involved in the biosynthesis of sterols and are considered major targets of azole antifungal agents. Here, we report on the expression and characterization of the MGL_0310 gene product...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.07.002
更新日期：2015-10-01 00:00:00
abstract：:Interferons (IFNs) are a family of pleiotropic cytokines used for the treatment of various viral infections and cancers. The low-cost production of IFNs with high biological value and the discovery of IFNs with improved properties are important for the treatment of these diseases as well as for understanding the physi...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(03)00187-6
更新日期：2003-10-01 00:00:00
abstract：:Darbepoetin alfa is an engineered and hyperglycosylated analog of recombinant human erythropoietin (EPO) which is used as a drug in treating anemia in patients with chronic kidney failure and cancer. This study desribes the secretory expression of a codon-optimized recombinant form of darbepoetin alfa in Leishmania ta...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.10.013
更新日期：2016-02-01 00:00:00
abstract：:A highly efficient cell-free translation system has been combined with suppressor tRNA technology to substitute nor-Tyr and 3-fluoro-Tyr in place of Tyr183 at the DNA polymerase active site of p66 of human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT). Supplementing the wild-type HIV-1 p51 RT subunit ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.07.019
更新日期：2004-11-01 00:00:00
abstract：:A novel and rapid procedure for the isolation of catalase from mouse liver, after prior treatment with the peroxisome proliferator perfluorooctanoic acid was developed using immobilized metal ion affinity chromatography involving chelation with zinc ions. The purification developed is simple, rapid (requiring 3 hours ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1997.0827
更新日期：1998-03-01 00:00:00
abstract：:Insoluble expression of intrinsically soluble proteins with native activity is potentially a promising alternative to soluble expression of folded protein or insoluble expression of unfolded protein requiring refolding. Here, we attempted to express highly soluble halophilic His-rich metal binding protein (HP) as inso...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2019.01.001
更新日期：2019-04-01 00:00:00
abstract：:Protein expression using the secretory pathway in Saccharomyces cerevisiae can lead to high amounts of overexpressed and secreted proteins in culture supernatants in a short period of time. These post-translational modified expression products can be purified up to >90% in a single step. The overexpression and secreti...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2000.1337
更新日期：2001-02-01 00:00:00
abstract：:Vascular leak syndrome (VLS) is the major dose-limiting toxicity of immunotoxin therapy. In our previous study, a modified PE38KDEL, denoted PE38KDELKQK, was engineered to eliminate VLS. The PE38KDELKQK-based immunotoxin has been proved to retain potent anti-tumor activity but with a remarkable attenuation in VLS. In ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2007.09.009
更新日期：2008-03-01 00:00:00
abstract：:Recombinant human interleukin-5 (rhIL-5) was expressed in baculovirus-infected insect cells and purified to homogeneity from the culture medium in a single chromatographic step. Beginning with a cDNA encoding the full-length precursor form of human IL-5, including the authentic secretory leader sequence, recombinant b...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1995.1009
更新日期：1995-02-01 00:00:00
abstract：:Fatty acid desaturases are enzymes that introduce double bonds into fatty acyl chains, among which stearoyl-acyl carrier protein desaturase (S-ACP-DES) was widely distributed in the plant kingdom. We cloned the cDNA coding for fab2/ssi2, an S-ACP-DES from Arabidopsis thaliana, into the vector pET30a and heterologously...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2009.08.011
更新日期：2010-02-01 00:00:00
abstract：:Enteroaggregative Escherichia coli (EAEC) heat-stable enterotoxin 1 (EAST1) is a 4.1k Da protein originally discovered in EAEC but known to be scattered in other diarrheagenic E. coli as well, possibly causing diarrhea in humans and animals. We report for the first time a method to express and purify EAST1 using the G...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2003.09.008
更新日期：2004-02-01 00:00:00
abstract：:Thymosin α1-thymopentin (Tα1-TP5) fusion peptide has been proved to be an immune regulator based on its higher immunoregulatory activity than Tα1 and TP5. To obtain Tα1-TP5 more effectively and economically, Tα1-TP5 was genetically fused to a self-cleaving intein-chitin binding domain tag for purification via chitin b...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2012.04.013
更新日期：2012-07-01 00:00:00
abstract：:Endo-1,4-β-mannanase is an enzyme that can catalyze the random hydrolysis of β-1,4-mannosidic linkages in the main chain of mannans, glucomannans and galactomannans and offers many applications in different biotechnology industries. Purification and kinetic properties of the endo-1,4-β-mannanase from recombinant Esche...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2018.01.009
更新日期：2018-06-01 00:00:00
abstract：:Huperzine A (HupA) is a drug used for the treatment of Alzheimer's disease. However, the biosynthesis of this medicinally important compound is not well understood. The HupA biosynthetic pathway is thought to be initiated by the decarboxylation of lysine to form cadaverine, which is then converted to 5-aminopentanal b...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2016.07.013
更新日期：2016-12-01 00:00:00
abstract：:We examined expression of two plant genes encoding coclaurine N-methyltransferase (CMT) and norcoclaurine synthase (NCS) in Escherichia coli from the Salmonella entericaprpBCDE promoter (P(prpB)) and compared it to that from the strongest IPTG-inducible promoter, P(T7). In contrast to our previous study showing slight...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.06.008
更新日期：2008-10-01 00:00:00
abstract：:Pseudomonas fluorescens is a robust protein expression system that is very well suited for high throughput protein expression for structural genomics studies. Since NMR spectroscopy and X-ray crystallography are both used by various investigators in structure elucidation studies, the availability of target proteins la...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.12.012
更新日期：2009-05-01 00:00:00
abstract：:Production of antibody fragments in heterologous hosts such as Escherichiacoli provides a unique and cost-effective method to develop engineered vectors for tumor targeting. A single-chain Fragment variable (scFv) of the murine monoclonal antibody MAb-B43.13 targeting CA125 in epithelial ovarian cancer was previously ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.07.007
更新日期：2014-10-01 00:00:00
abstract：:2-Deoxy-d-ribose-5-phosphate aldolase (DERA) catalyzes the aldol reaction between two aldehydes and is thought to be a potential biocatalyst for the production of a variety of stereo-specific materials. A gene encoding DERA from the extreme halophilic archaeon, Haloarcula japonica, was overexpressed in Escherichia col...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2016.05.009
更新日期：2016-10-01 00:00:00
abstract：:To investigate the expression and purification of an unstable heterologous protein in Pichia pastoris, the cDNA of H5-lysozyme, a hen egg lysozyme mutant with a hydrophobic pentapeptide (Phe-Phe-Val-Ala-Pro) fused to the carboxyl terminus, was integrated into the genome of P. pastoris. It was found that medium composi...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(02)00642-3
更新日期：2003-02-01 00:00:00
abstract：:A method was developed for the affinity purification of human complement properdin. The preparation is part of an integrated scheme in which over 20 human plasma proteins can be recovered in a highly purified form. The yield of properdin was 5.9 mg from 3 liters of plasma, amounting to a 28% recovery. The crucial step...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1994.1026
更新日期：1994-04-01 00:00:00
abstract：:Hsp47 is regarded as a collagen-specific chaperone with several suggested roles in collagen biosynthesis under normal and disease conditions. We describe here a procedure for the expression and purification of Hsp47 in Escherichia coli using the IMPACT expression system (New England Biolabs) where the guest gene is fu...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2001.1470
更新日期：2001-10-01 00:00:00
