Abstract:
:Here we present a biophysical, structural, and computational analysis of the directed evolution of the human DNA repair protein O(6)-alkylguanine-DNA alkyltransferase (hAGT) into SNAP-tag, a self-labeling protein tag. Evolution of hAGT led not only to increased protein activity but also to higher stability, especially of the alkylated protein, suggesting that the reactivity of the suicide enzyme can be influenced by stabilizing the product of the irreversible reaction. Whereas wild-type hAGT is rapidly degraded in cells after alkyl transfer, the high stability of benzylated SNAP-tag prevents proteolytic degradation. Our data indicate that the intrinstic stability of a key α helix is an important factor in triggering the unfolding and degradation of wild-type hAGT upon alkyl transfer, providing new insights into the structure-function relationship of the DNA repair protein.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Mollwitz B,Brunk E,Schmitt S,Pojer F,Bannwarth M,Schiltz M,Rothlisberger U,Johnsson Kdoi
10.1021/bi2016537subject
Has Abstractpub_date
2012-02-07 00:00:00pages
986-94issue
5eissn
0006-2960issn
1520-4995journal_volume
51pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::Hepatic mixed-function oxidase metabolism of the ubiquitous pollutant polychlorinated biphenyls (PCBs) is implicated in their toxification and detoxification. We used dichlorobiphenyls (DCBs) as models to investigate the effect of the chloro substituent sites on this metabolism experimentally and by molecular orbital ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00529a009
更新日期:1981-12-22 00:00:00
abstract::Seven hydrophobic residues ranging in size from glycine to phenylalanine have been substituted for the wild-type methionine residue at position 13 in a 15-residue truncated version (S15) of S-peptide, the small component of ribonuclease S. Complexes of both S-15 and the seven variants with S-protein yielded isomorphou...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00164a005
更新日期:1992-12-15 00:00:00
abstract::Single-tryptophan-containing mutants of low adenylation state Escherichia coli glutamine synthetase (wild type has two tryptophans at positions 57 and 158) have been constructed and studied by multifrequency phase/modulation fluorescence spectroscopy. The W57L mutant (retains tryptophan at residue 158) and the W158S m...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00228a008
更新日期:1991-04-09 00:00:00
abstract::ATP-dependent Mg2+ accumulation in isolated mitochondria occurs predominantly in the matrix and inner membrane compartments. In mitochondria contaminated with lysosomes, the time course and magnitude of ATP-dependent Mg2+ accumulation are influenced by various cytoplasmic substances, besides substrates of the citric a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00656a013
更新日期:1976-06-01 00:00:00
abstract::Protein disulfide-isomerase, a protein localized to the lumen of the endoplasmic reticulum of eukaryotic cells, catalyzes the posttranslational formation and rearrangement of protein disulfide bonds. As isolated from bovine liver, the enzyme contains 0.8 free sulfhydryl group per mole of protein monomer and 3.1 disulf...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00444a023
更新日期:1989-09-05 00:00:00
abstract::The binding of the R17 coat protein to synthetic RNAs containing one or two coat protein binding sites was characterized by using nitrocellulose filter and gel-retention assays. RNAs with two available sites bound coat protein in a cooperative manner, resulting in a higher affinity and reduced sensitivity to pH, ionic...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00502a006
更新日期:1990-12-18 00:00:00
abstract::The thermodynamics of the interactions of serum apotransferrin (T) and holotransferrin (TFe(2)) with ceruloplasmin (Cp), as well as those of human lactoferrin (Lf), were assessed by fluorescence emission spectroscopy. Cp interacts with two Lf molecules. The first interaction depends on pH and μ, whereas the second doe...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1014503
更新日期:2010-12-07 00:00:00
abstract::Sir2 protein deacetylases (or sirtuins) catalyze NAD+-dependent conversion of epsilon-amino-acetylated lysine residues to deacetylated lysine, nicotinamide, and 2'-O-acetyl-ADP-ribose. Small-molecule modulation of sirtuin activity might treat age-associated diseases, such as type II diabetes, obesity, and neurodegener...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi7013294
更新日期:2007-12-18 00:00:00
abstract::Phytochromes are widespread red/far-red photosensory proteins well known as critical regulators of photomorphogenesis in plants. It is often assumed that natural selection would have optimized the light sensing efficiency of phytochromes to minimize nonproductive photochemical deexcitation pathways. Surprisingly, the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi500108s
更新日期:2014-05-06 00:00:00
abstract::PsbU is a lumenal peripheral protein in the photosystem II (PS II) complex of cyanobacteria and red algae. It is thought that PsbU is replaced functionally by PsbP or PsbQ in plant chloroplasts. After the discovery of PsbP and PsbQ homologues in cyanobacterial PS II [Thornton et al. (2004) Plant Cell 16, 2164-2175], w...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi047539k
更新日期:2005-09-13 00:00:00
abstract::The rate of deuterium exchange of a purine 8CH group in DNA is highly sensitive to both macromolecular secondary structure and intermolecular interactions which restrict solvent access to the major groove [Lamba, O.P., Becka, R., & Thomas, G.J., Jr. (1990) Biopolymers 29, 1465-1477]. We have exploited the sensitivity ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00127a012
更新日期:1992-03-31 00:00:00
abstract::Raman spectroscopy was used to study the phase behavior of several semisynthetic species of the acidic glycosphingolipid cerebroside sulfate (CBS) which occur in myelin. The C-H stretching mode region at 2800-3100 cm-1 of C18:0-CBS, C24:0-CBS, and C26:0-CBS, and the alpha-hydroxy fatty acid species C18:0h-CBS, was stu...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00121a041
更新日期:1992-02-18 00:00:00
abstract::We examined the metabolism of exogenously added 13C-labeled fructose 1,6-bisphosphate (either labeled at the first and sixth carbons or labeled at the first carbon only) and of [2-13C]glucose in well-oxygenated and well-superfused hog carotid artery segments. Exogenously added fructose 1,6-bisphosphate was utilized by...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00004a027
更新日期:1995-01-31 00:00:00
abstract::X-ray solution scattering data from skeletal muscle troponin C and from calmodulin have been measured. Modeling studies based on the crystal structure coordinates for these proteins show discrepancies between the solution data and the crystal structure that indicate that if the size and shape of the globular domains a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00403a011
更新日期:1988-02-09 00:00:00
abstract::The complex of ribonuclease A (RNase A) with uridine vanadate (U-V), a transition-state analogue, has been studied with 51V and proton NMR spectroscopy in solution and by neutron diffraction in the crystalline state. Upon the addition of aliquots of U-V at pH 6.6, the C epsilon-H resonances of the two active-site hist...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00329a038
更新日期:1985-04-09 00:00:00
abstract::The PEGylated hemoglobin (Hb) has been evaluated as a potential blood substitute. In an attempt to understand the autoxidation of the PEGylated Hb, we have studied the autoxidation of the PEGylated Hb site-specifically modified at Cys-93(beta) or at Val-1(beta). PEGylation of Hb at Cys-93(beta) perturbed the heme envi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi800906z
更新日期:2008-10-14 00:00:00
abstract::The search for a rational method of developing an antimalarial vaccine (malaria caused by Plasmodium falciparum) consists of blocking receptor-ligand interaction. Conserved peptides derived from proteins involved in invasion and having strong red blood cell binding ability have thus been identified; immunization studi...
journal_title:Biochemistry
pub_type: 历史文章,杂志文章
doi:10.1021/bi050214z
更新日期:2005-05-10 00:00:00
abstract::A high-resolution solution conformation of a novel conotoxin, [Pro 7,13] alpha A-conotoxin PIVA, GCCGSYPNAACHPCSCKDROSYCGQ-NH2, has been determined by two-dimensional 1H NMR methods and distance geometry calculations. The total of 324 NOE-derived interproton distance restraints including 33 long-range NOE restraints a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi962301k
更新日期:1997-02-18 00:00:00
abstract::Amyloid beta peptide (A beta) is released into the media of a variety of cells in culture during normal metabolism. The discovery of several missense mutations within or flanking the A beta region of the beta amyloid precursor protein (beta APP) in familial Alzheimer's disease provides strong evidence for a role of al...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00181a016
更新日期:1994-04-19 00:00:00
abstract::To better understand origin recognition and initiation of DNA replication, we have examined by NMR complexes formed between the origin-binding domain of SV40 T antigen (T-ag-obd), the initiator protein of the SV40 virus, and cognate and noncognate DNA oligomers. The results reveal two structural effects associated wit...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi030228+
更新日期:2004-06-08 00:00:00
abstract::RP2 is a ubiquitously expressed protein encoded by a gene associated with X-linked retinitis pigmentosa (XLRP), a retinal degenerative disease that causes severe vision loss. Previous in vitro studies have shown that RP2 binds to ADP ribosylation factor-like 3 (Arl3) and activates its intrinsic GTPase activity, but th...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1005249
更新日期:2010-09-07 00:00:00
abstract::Complete resonance assignments of nonexchangeable base protons and sugar protons in d-GGTACGCGTACC at 500 MHz have been obtained by two-dimensional correlated spectroscopy (COSY) and nuclear Overhauser enhancement spectroscopy (NOESY). The characteristic phase-sensitive multiplet patterns of the ntrasugar cross peaks ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00410a050
更新日期:1988-05-17 00:00:00
abstract::The structures of beta class carbonic anhydrases (beta-CAs) determined so far fall into two distinct subclasses based on the observed coordination of the catalytic zinc (Zn2+) ion. The subclass of beta-CAs that coordinate Zn2+ tetrahedrally with four protein-derived ligands is represented by the structures of ortholog...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi052272q
更新日期:2006-04-11 00:00:00
abstract::In a family of proteins, often the three-dimensional structure has been experimentally determined only for one member or a few members of the family. Homology modeling can be used to model the structures of all other members of the family and thus allow comparison of these structures. This approach was applied to hete...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi026729x
更新日期:2002-12-10 00:00:00
abstract::Sulfite oxidase (SO) catalyzes the physiologically critical conversion of sulfite to sulfate. Enzymatic activity is dependent on the presence of the metal molybdenum complexed with a pyranopterin-dithiolene cofactor termed molybdopterin. Comparison of the amino acid sequences of SOs from a variety of sources has ident...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1001954
更新日期:2010-05-11 00:00:00
abstract::In globular proteins, there are intermolecular hydrogen bonds between protein and water molecules, and between water molecules, which are bound with the proteins, in addition to intramolecular hydrogen bonds. To estimate the contribution of these hydrogen bonds to the conformational stability of a protein, the thermod...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9910169
更新日期:1999-09-28 00:00:00
abstract::To test whether the Y-shaped conformation of the hammerhead ribozyme is maintained throughout the catalytic pathway, the cleavage properties of circular substrates which bind the ribozyme through helices I and II were determined. Constraining the position of helices I and II in this manner did not significantly alter ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi980307z
更新日期:1998-06-30 00:00:00
abstract::The UV endonucleases [endodeoxyribonuclease (pyrimidine dimer), EC 3.1.25.1] from Micrococcus luteus and bacteriophage T4 possess two catalytic activities specific for the site of cyclobutane pyrimidine dimers in UV-irradiated DNA: a DNA glycosylase that cleaves the 5'-glycosyl bond of the dimerized pyrimidines and an...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00386a011
更新日期:1987-06-16 00:00:00
abstract::Polymerase eta is a member of the Y family of DNA polymerases which is able to bypass thymine dimers efficiently and in a relatively error-free manner. To elucidate the mechanism of dimer bypass, the efficiency of dAMP and pyrene nucleotide insertion opposite the thymine dimer and its N3-methyl derivatives was determi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0345687
更新日期:2003-08-12 00:00:00
abstract::We report here that the DNA strand scission by dynemicin A is not only sequence-specific but also conformation-specific. The salt-induced B----Z conformational transition dramatically enhanced the cleavage by dynemicin A in a B-Z junction region. By contrast, the bleomycin-Fe(II) complex, the elsamicin A-Fe(II) comple...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00144a019
更新日期:1992-07-28 00:00:00