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Inteins (0):
Recent article
Inteins
Abstract
Protein splicing is a post-translational process by which an intervening polypeptide, the intein, excises itself from the flanking polypeptides, the exteins, coupled to ligation of the exteins. The lon protease of Pyrococcus abyssi (Pab) is interrupted by an intein. When over-expressed as a fusion p...
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PMID: 21094142
PDF is available here.
PDF is available here.
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Protein Science (19)8 2010
Abstract
We previously proposed that Class 3 inteins splice by a variation in the standard intein-mediated protein splicing mechanism that includes a novel initiating step leading to the formation of a previously unrecognized branched intermediate. In this mechanism defined with the Class 3 prototypic Mycoba...
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PMID: 20521254
PDF is available here.
PDF is available here.
Journal of Molecular Biology (400)4 2010
Abstract
We have studied the conformations and dynamics of the minimized and engineered intein DeltaDeltaIhh-V67CM and a single V67L mutant, DeltaDeltaIhh-L67CM, by solution NMR. Chemical shift perturbations established that the V67L mutation causes global changes, including changes at the N-terminus and C-t...
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PMID: 20562025
PDF is available here.
PDF is available here.
Abstract
A mutant of green fluorescent protein (GFPmut3*) from the jellyfish Aequorea victoria was cyclized in vitro and in vivo by the use of a naturally split intein from the dnaE gene of Synechocystis species PCC6803 (Ssp). Cyclization of GFPmut3* was confirmed by amino acid sequencing and resulted in an...
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PMID: 20372012
PDF is available here.
PDF is available here.
Abstract
Among human antimicrobial peptides (hAMPs), DCD-1L has a broad spectrum of antimicrobial activity over a wide pH range and in high salt concentrations. It offers a promising alternative to conventional antibiotics. The 458-bp-long dermcidin cDNA was amplified by PCR using a human fetal cDNA library...
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PMID: 20208440
PDF is available here.
PDF is available here.
Abstract
Eight of fifty Iranian AT patients (16%) exhibited a C > T transition in exon 2 (c342C > T) of the BAK gene, while none of the healthy controls had such alteration (P = 0.0001). Higher frequency of another nucleotide substitution in the noncoding region of exon 7 in BAX gene (6855G > A) was also ide...
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PMID: 19898928
PDF is available here.
PDF is available here.
Protein & Peptide Letters (17)10 2010
Abstract
To facilitate expression and purification of an analog of GLP-1 (mGLP-1), an intein system was employed in this study. A recombinant fusion protein, CBD-DnaB-mGLP-1, was constructed and expressed in the form of inclusion body. After refolding, the intein-mediated self-cleavage was triggered by pH an...
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PMID: 20441562
PDF is available here.
PDF is available here.
Acta Microbiologica Sinica (49)12 2009
Abstract
Through IPTG induction for expression of recombinant protein it displayed an obvious protein band as predicted size of ABCA1 on SDS-PAGE gel. Western blotting using His-Tag specific antibody confirmed that this protein band is trans-spliced ABCA1. CONCLUSION: The data demonstrated that Ssp DnaE inte...
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PMID: 20222445
PDF is available here.
PDF is available here.
Abstract
We describe a simple yet widely applicable two-color site-specific protein labeling method utilizing Npu DnaE split intein....
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PMID: 19645470
PDF is available here.
PDF is available here.
Abstract
We have combined NMR pK(a) determination with quantum mechanics/molecular mechanics (QM/MM) modeling to study engineered inteins from Mycobacterium tuberculosis (Mtu) RecA intein. We demonstrate a dramatic pK(a) shift for the invariant B-block histidine, the most conserved residue among inteins. The...
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PMID: 19630416
PDF is available here.
PDF is available here.
Abstract
Apolipoprotein E (apoE) is an exchangeable apolipoprotein that functions as a ligand for members of the LDL receptor family, promoting lipoprotein clearance from the circulation. Productive receptor binding requires that apoE adopt an LDL receptor-active conformation through lipid association, and s...
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PMID: 19098282
PDF is available here.
PDF is available here.
B M B Reports (42)6 2009
Abstract
We have examined human-mouse homologous intronless disease and non-disease genes alongside their extent of sequence conservation, tissue expression, domain and gene ontology composition to get an idea regarding evolutionary and functional attributes. We show that selection has significantly discrimi...
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PMID: 19558794
PDF is available here.
PDF is available here.
Abstract
Inteins or 'internal proteins' are coding sequences that are transcribed and translated with flanking sequences (exteins). After translation, the inteins are excised by an autocatalytic process and the host protein assumes its normal conformation and develops its expected function. These parasitic g...
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PMID: 19547879
PDF is available here.
PDF is available here.
Nucleic Acids Research (37)8 2009
Abstract
Inteins are genetic elements, inserted in-frame into protein-coding genes, whose products catalyze their removal from the protein precursor via a protein-splicing reaction. Intein domains can be split into two fragments and still ligate their flanks by a trans-protein-splicing reaction. A bioinforma...
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PMID: 19264795
PDF is available here.
PDF is available here.
Abstract
We report a new family of bacterial intein-like domains (BILs) identified in ten proteins of four diverse predatory bacteria. BILs belong to the HINT (Hedgehog/Intein) superfamily of domains that post-translationally self-process their protein molecules by protein splicing and self-cleavage. The new...
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PMID: 19153786
PDF is available here.
PDF is available here.
Methods in Molecular Biology (532)532 2009
Abstract
This chapter defines the agents that provide for the movement of genetic material which fuels the adaptive potential of life on our planet. The chapter has been structured to be broadly comprehensive, arbitrarily categorizing the mobilome into four classes: (1) transposons, (2) plasmids, (3) bacteri...
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PMID: 19271177
PDF is available here.
PDF is available here.
Abstract
We have identified inteins in the highly conserved 3.2 region of the PRP8 protein from species of the genus Penicillium. These inteins are integrated at the same position as that which has recently been described in PRP8 proteins from different strains of Cryptococcus neoformans and several ascomyce...
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PMID: 19253333
PDF is available here.
PDF is available here.
Abstract
Twenty years ago, evidence that one gene produces two enzymes via protein splicing emerged from structural and expression studies of the VMA1 gene in Saccharomyces cerevisiae. VMA1 consists of a single open reading frame and contains two independent genetic information for Vma1p (a catalytic 70-kDa...
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PMID: 19907126
PDF is available here.
PDF is available here.
BMC Biotechnology (9)1 2009
Abstract
Targeting to the periplasm of E. coli allowed efficient production of correctly-folded single-domain antibody (sdAb)-intein fusions proteins. On column purification and 2-mercapthoethanesulfonic acid (MESNA)-induced cleavage yielded single-domain antibodies with a reactive C-terminal MESNA thioester...
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PMID: 19619333
PDF is available here.
PDF is available here.
Abstract
We show a new reporter gene assay for detecting Ras-Raf-1 interactions based on protein splicing of transcription factors with DnaE inteins. The protein splicing induces connection of a DNA-binding protein (modified LexA; mLexA) with a transcription activation domain of a herpes simplex virus protei...
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PMID: 19685309
PDF is available here.
PDF is available here.
Abstract
A method has been developed that eliminates the need for complex chromatographic apparatus in the purification of recombinant proteins expressed in Escherichia coli. This method is similar to conventional affinity-tag separations, but the affinity resin is replaced by polyhydroxybutyrate (PHB) parti...
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PMID: 18988026
PDF is available here.
PDF is available here.
ChemBioChem (9)18 2008
Segmental isotopic labelling of a multidomain protein by protein ligation by protein trans-splicing.
Abstract
Abstract
To produce recombinant Maxadilan using gene engineering technology, the gene of recombinant Maxadilan which expressed in protocaryon were designed and synthesized according to the amino acid sequences of Maxadilan. The recombinant plasmid pKYB-MAX was constructed and transformed into host bacteria E...
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PMID: 19306574
PDF is available here.
PDF is available here.
Abstract
We report here a protease sensing nanoplatform based on semiconductor nanocrystals or quantum dots (QDs) and bioluminescence resonance energy transfer (QD-BRET) to detect the protease activity in complex biological samples. These nanosensors consist of bioluminescent proteins as the BRET donor, quan...
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PMID: 18922019
PDF is available here.
PDF is available here.
Biochimica et Biophysica Acta (1784)11 2008
Abstract
A strategy for simultaneous purification and refolding of proteins overexpressed with an intein tag is described. A recombinant lipase overexpressed in Escherichia coli ER2566 with the intein tag and obtained as inclusion bodies was solubilized in buffer containing 8 M urea or cetyltrimethylammonium...
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PMID: 18725331
PDF is available here.
PDF is available here.
Abstract
A library with potential to produce six amino acids cyclic peptides was prepared using pET-28a as the starting plasmid. pVmut was used to amplify the Int(C)-dnaB-N-Int(N) fragment that was inserted into pET28a to give pEV. On pEV, DnaB split intein was expressed under the strong T7 promoter. Analyse...
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PMID: 19256340
PDF is available here.
PDF is available here.
Analytical Biochemistry (381)1 2008
Abstract
Multiple recombinant proteins can be expressed simultaneously by inoculating multiple seed cultures into a single growth medium and inducing protein expression at a single time point. Up to three recombinant proteins can be individually purified from such a mixed culture (cocultivation) through the...
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PMID: 18602361
PDF is available here.
PDF is available here.
Abstract
We have combined Invitrogen's Gateway cloning technology with self-cleaving purification tags to generate a new system for rapid production of recombinant protein products. To accomplish this, we engineered our previously reported DeltaI-CM cleaving intein to include a Gateway cloning recognition se...
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PMID: 18727029
PDF is available here.
PDF is available here.
ChemBioChem (9)14 2008
Abstract
We have recently reported a new approach in which a short cysteine-containing tag (CysTag) fused to one part of a split intein is first modified with a sulfhydryl-reactive probe. In a second step, protein trans-splicing is used to link the labelled CysTag to a target protein that has been expressed...
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PMID: 18756552
PDF is available here.
PDF is available here.
Journal of Biological Chemistry (283)37 2008
Abstract
We recently reported an artificially split intein based on the Ssp DnaB mini-intein that consists of a synthetic N-terminal intein fragment (Int(N)) and a recombinant C-terminal part (Int(C)), which are 11 and 143 amino acids in length, respectively. This intein holds great promise for the preparati...
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PMID: 18625708
PDF is available here.
PDF is available here.
Abstract
We have identified 69 putative open reading frames, including three apparent genes (thymidylate synthase, terminase, and RNR alpha subunit genes) that are interrupted by an intein. Protein-splicing activities of these inteins were demonstrated in Escherichia coli. Three structural proteins including...
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PMID: 18599103
PDF is available here.
PDF is available here.
Journal of Bacteriology (190)17 2008
Abstract
We describe the ability of the intron-encoded RNAs (ribozymes) to retard bacterial growth rate (toxicity) and examine the functionality and phylogenetic history of Cbu.DnaB. When expressed in Escherichia coli, both introns repressed growth, with Cbu.L1917 being more inhibitory. Both ribozymes were f...
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PMID: 18606739
PDF is available here.
PDF is available here.
Abstract
A recent species status investigation of the pathogenic fungus Paracoccidioides brasiliensis suggested the existence of three cryptic species. In the present study, the sequences of the PRP8 intein from P. brasiliensis isolates belonging to the three described genetic groups and two unidentified iso...
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PMID: 18672080
PDF is available here.
PDF is available here.
Abstract
Chemical-enzymatic synthesis of human Epidermal Growth Factor (hEGF) cDNA has been performed, following by cloning into expression vector pTWIN1 (New England Biolabs). The resulting recombinant fusion protein expressed in Escherichia coli consisted of the N-terminal chitin-binding domain, mini-intei...
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PMID: 18599312
PDF is available here.
PDF is available here.
Journal of Microbiology (46)4 2008
Abstract
These results suggest that the novel reporter system can be exploited to develop new inteins with high efficiency of in vivo protein cyclization....
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PMID: 18758738
PDF is available here.
PDF is available here.
Abstract
We investigate the effect of Ca(2+), Mn(2+), Ni(2+), Zn(2+), Cd(2+), Pb(2+), and [Co(NH(3))(6)](3+) on the first step of splicing of the Saccharomyces cerevisiae mitochondrial group II intron Sc.ai5gamma. We find that this group II intron is very sensitive to the presence of divalent metal ions othe...
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PMID: 18528718
PDF is available here.
PDF is available here.
Journal of Chromatography A (1194)2 2008
Abstract
A simple, low-cost, and scalable protein purification method was developed by using a biodegradable regenerated amorphous cellulose (RAC) with a binding capacity of up to 365 mg protein per gram of RAC. The recombinant protein with a cellulose-binding module (CBM) tag can be specifically adsorbed by...
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PMID: 18468611
PDF is available here.
PDF is available here.
Abstract
The high frequency and urge UI was significantly associated with a lower level of physical fitness and self-rated health. Our findings suggest that active lifestyle, which was focused on a decreased BMI and increased walking ability, may contribute to the prevention of UI....
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PMID: 18622117
PDF is available here.
PDF is available here.
Abstract
The low yield and poor folding efficiency in vivo of soluble and active recombinant cysteine-rich proteins expressed in Escherichia coli are a major challenge for large-scale protein production and purification. Expression vectors containing Buthus martensii Karsch insect toxin (BmK IT) fused to the...
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PMID: 18289876
PDF is available here.
PDF is available here.
Abstract
We found a new schizophrenia risk and protective haplotypes in intron VII of DTNBP1; one of the most important candidate genes for this disorder, to-date. However, no association was found between DAO, DAOA, NRG1 and RGS4 and schizophrenia. The hypothesis that gene-gene interaction in these five gen...
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PMID: 17408693
PDF is available here.
PDF is available here.
Abstract
These findings strengthen the support for an association between PSD and lower-expressing alleles of 5-HTTLPR....
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PMID: 18441666
PDF is available here.
PDF is available here.
Abstract
Inteins are internal protein splicing elements that can autocatalytically self-excise from their host protein and ligate the protein flanks (exteins) with a peptide bond. Large inteins comprise independent protein splicing and endonuclease domains whereas mini-inteins lack the central endonuclease d...
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PMID: 18054328
PDF is available here.
PDF is available here.
Abstract
As a novel attempt for the intracellular recombinant protein over expression and easy purification from Pichia pastoris, the therapeutic cytokine human granulocyte macrophage colony stimulating factor (hGMCSF) gene was fused to an intein-chitin-binding domain (gene from pTYB11 vector) fusion tag by...
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PMID: 18309571
PDF is available here.
PDF is available here.
Abstract
We tried to express ABP-CM4 in Escherichia coli cells using either the GST fusion system or the intein-mediated fusion expression system. In order to investigate the possible use of these two fusion partners in cationic small peptide expression and purification, a mutant ABP-CMt, which is a highly p...
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PMID: 17977015
PDF is available here.
PDF is available here.
Abstract
We followed the formation of these adducts as well as ways to avoid them with electrospray ionization mass spectrometry using one of our target proteins, Triticum aestivum (wheat) E(c)-1, a plant metallothionein with the ability to bind a total of six zinc or cadmium ions in the form of metal-thiola...
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PMID: 18032065
PDF is available here.
PDF is available here.
Developmental Biology (313)2 2008
Abstract
We have identified five intein-containing genes in the anthozoan cnidarian Nematostella vectensis, two of which (NvHh1 and NvHh2) contain definitive hedgehog ligand domains, suggesting that to date, cnidarians are the earliest branching metazoan phylum to possess definitive Hh orthologs. Expression...
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PMID: 18068698
PDF is available here.
PDF is available here.
Abstract
One of the most important problems in biotechnology today is development of simple and cheap techniques for protein purification. This is why inteins and protein splicing phenomena are of a great interest for protein purification. Modification of inteins by affinity tags permits to use general metho...
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PMID: 19140330
PDF is available here.
PDF is available here.
Abstract
Abstract
Abstract
We cloned twelve additional subfamily Ialpha enzymes from an evolutionarily divergent set of organisms. Nine were purified to homogeneity after heterologous expression in Escherichia coli in native, intein-tagged or His(6)-tagged forms. The two Saccharomyces cerevisiae isoforms were recombinantly pr...
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PMID: 17964807
PDF is available here.
PDF is available here.
Abstract
We will discuss several valuable tags for addressing this problem, and comment on the potential usage of tagged fusion protein for molecular imaging....
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PMID: 17981663
PDF is available here.
PDF is available here.
Abstract
A two-intein purification system was developed for the affinity purification of GFPmut3*, a mutant of green fluorescent protein. The GFPmut3* was sandwiched between two self-cleaving inteins. This approach avoided the loss of the target protein which may result from in vivo cleavage of a single inte...
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PMID: 17973109
PDF is available here.
PDF is available here.
Construction of a small-molecule-integrated semisynthetic split intein for in vivo protein ligation.
Abstract
A new split intein-based protein ligation tool that is synthetically accessible and can be used for protein semisynthesis on the cell surface and potentially inside cells has been constructed.
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PMID: 18049731
PDF is available here.
PDF is available here.
Analytical Biochemistry (371)2 2007
Abstract
We describe a robust fluorescence intensity assay for monitoring the enzymatic activity of deubiquitinating proteases, which reverse ubiquitin modifications and comprise over 100 members in humans. The assay was developed for the catalytic domain of human ubiquitin-specific protease 2 (USP2) and hum...
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PMID: 17869210
PDF is available here.
PDF is available here.
Abstract
We found that in some cases, fusion to annexin B1 can promote the solubility of heterologous proteins. The production of soluble and highly active of interleukin-2 and low-molecular single-chain urokinase in our results proved that the system was a novel, cheap and effective fusion expression system...
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PMID: 17768617
PDF is available here.
PDF is available here.
Abstract
To produce recombinant Buthus martensii Karsch insect toxin (BmK IT), BmK IT cDNA which fused a hexahistidine sequence at the C-terminus by PCR was inserted into pTWIN1 expression vector fused in frame with an upstream Ssp DnaB intein gene. The expression plasmid was transformed into E. coli BL21 (D...
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PMID: 18257224
PDF is available here.
PDF is available here.
Abstract
We have developed a new method to generate AIP structures using an engineered DnaB mini-intein from Synechocystis sp. strain PCC6803. In the method, an oligonucleotide encoding the AIP is ligated to the intein and the fusion protein is expressed and purified by affinity chromatography. To produce th...
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PMID: 17693565
PDF is available here.
PDF is available here.
Abstract
We sequenced the coding exons of the cardiac troponins T (TNNT2) and I (TNNI3) genes in 115 Spanish HCM-patients (32% with a family history of the disease). Only two (2%) had mutations in the TNNT2 (Arg278>Cys and Arg92>Lys). These mutations were associated with variable clinical outcomes. No patien...
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PMID: 17101185
PDF is available here.
PDF is available here.
Biological Chemistry (388)9 2007
Abstract
I molecules for presentation to the immune system on the cell surface. MHC class I molecules are highly selective and specific in terms of ligand binding. Variability of peptides produced in living cells arises in a variety of ways, ensuring fast and efficient immune responses. Substitution of const...
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PMID: 17696779
PDF is available here.
PDF is available here.
Biochimica et Biophysica Acta (1774)8 2007
Abstract
As the Cne PRP8 intein is active and exists in an essential gene of an important fungal pathogen, inhibitors of splicing and assays for intein activity are of interest. The self-splicing activity of Cne PRP8, the intein from the Prp8 gene of Cryptococcus neoformans, was assessed in different heterol...
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PMID: 17604706
PDF is available here.
PDF is available here.