The design of an alternative, covalently flavinylated 6-hydroxy-D-nicotine oxidase by replacing the FAD-binding histidine by cysteine and reconstitution of the holoenzyme with 8-(methylsulfonyl)FAD.

Abstract:

:The cofactor of several flavoenzymes is autocatalytically bound to the polypeptide via a histidyl(N3)-(8alpha)-FAD linkage which makes the generation of apoenzyme difficult. We introduced an alternative covalent protein-FAD bond at the active site of 6-hydroxy-D-nicotine oxidase (6HDNO) by replacing the FAD-binding histidine with cysteine. The resulting mutant enzyme was expressed with noncovalently attached cofactor. Incubation with 8-(methylsulfonyl)FAD, and less efficiently with 8-chloro-FAD, resulted in the spontaneous replacement of the noncovalently bound FAD by the flavin derivative and the formation of an 8-(N-acetylcysteinyl)FAD linkage. The flavinylated 6HDNO.cys exhibited close to wild-type activity levels. This strategy may be generally applicable to the attachment of artificially designed flavin derivatives to the active site of covalently flavinylated enzymes.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Stoltz M,Henninger HP,Brandsch R

doi

10.1016/0014-5793(96)00438-3

subject

Has Abstract

pub_date

1996-05-20 00:00:00

pages

194-6

issue

2-3

eissn

0014-5793

issn

1873-3468

pii

0014-5793(96)00438-3

journal_volume

386

pub_type

杂志文章
  • Progranulin compensates for blocked IGF-1 signaling to promote myotube hypertrophy in C2C12 myoblasts via the PI3K/Akt/mTOR pathway.

    abstract::It is well known that growth hormone (GH)-induced IGF-1 signaling plays a dominant role in postnatal muscle growth. Our previous studies have identified a growth factor, progranulin (PGRN), that is co-induced with IGF-1 upon GH administration. This result prompted us to explore the function of PGRN and its association...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2012.07.077

    authors: Hu SY,Tai CC,Li YH,Wu JL

    更新日期:2012-09-21 00:00:00

  • Control region and gastric specific transcription of the rat H+,K(+)-ATPase alpha subunit gene.

    abstract::The rat gastric H+,K(+)-ATPase alpha subunit gene was cloned and the nucleotide sequence of its 5'-upstream region was determined. Sequence comparison with the corresponding part of the human gene indicated the presence of highly conserved regions which may be important for specific transcription of the alpha subunit ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(91)80404-q

    authors: Oshiman K,Motojima K,Mahmood S,Shimada A,Tamura S,Maeda M,Futai M

    更新日期:1991-04-09 00:00:00

  • The N intermediate of bacteriorhodopsin at low temperatures: stabilization and photoconversion.

    abstract::In aqueous suspensions of purple membranes (pH 10.2, 0.4 M KCl) an intermediate having an absorption maximum at 570-575 nm (at -196 degrees C) was produced by first heating the M intermediate up to -30 degrees C and then stabilizing it by subsequent cooling to -60 degrees C. We suggest that this species is the interme...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(90)80380-2

    authors: Balashov SP,Imasheva ES,Litvin FF,Lozier RH

    更新日期:1990-10-01 00:00:00

  • Regulation of human growth and differentiation factor 3 gene expression by NANOG in human embryonic carcinoma NCCIT cells.

    abstract::We investigated transactivation by NANOG in regulating growth and differentiation factor 3 (GDF3) expression in NCCIT cells. GDF3 expression was affected by shRNA-mediated downregulation and by exogenous overexpression of NANOG specifically, as well as by retinoic acid-mediated differentiation. GDF3 transcription was ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2012.08.013

    authors: Park SW,Lim HY,Do HJ,Sung B,Huh SH,Uhm SJ,Song H,Chung HJ,Kim JH,Kim NH,Kim JH

    更新日期:2012-09-21 00:00:00

  • GLUT4 translocation by insulin in intact muscle cells: detection by a fast and quantitative assay.

    abstract::We report a rapid and sensitive colorimetric approach to quantitate the amount of glucose transporters exposed at the surface of intact cells, using L6 muscle cells expressing GLUT4 containing an exofacial myc epitope. Unstimulated cells exposed to the surface 5 fmol GLUT4myc per mg protein. This value increased to 10...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(98)00423-2

    authors: Wang Q,Khayat Z,Kishi K,Ebina Y,Klip A

    更新日期:1998-05-08 00:00:00

  • Expression of the cell-adhesion molecule VCAM-1 by stromal cells is necessary for osteoclastogenesis.

    abstract::Osteoblastic cells have been shown to be involved in osteoclast formation through cell to cell contacts. This study was designed to examine the possible function of vascular cell adhesion molecule 1 (VCAM-1) during osteoclastogenesis. As a source for stromal cells we used the recently established mouse bone marrow str...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(96)01495-0

    authors: Feuerbach D,Feyen JH

    更新日期:1997-01-27 00:00:00

  • Rate limitation within a single enzyme is directly related to enzyme intermediate levels.

    abstract::The extents to which different rate constants limit the steady-state rate of an isolated enzyme can be quantified as the control coefficients of those constants and elemental steps. We have found that the sum of the control coefficients of rate constants characterising unidirectional rates depleting a particular enzym...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(94)00617-2

    authors: Kholodenko BN,Westerhoff HV,Brown GC

    更新日期:1994-07-25 00:00:00

  • An EVH1/WH1 domain as a key actor in TGFbeta signalling.

    abstract::EVH1 (enabled VASP (vasodilator-stimulated protein) homology 1)/WH1 (WASP (Wiskott-Aldrich syndrome protein) homology 1) domains, present in Ena VASP and WASP, are protein interaction modules specialised in binding proline-rich ligands. An EVH1/WH1 domain is here identified in the recently cloned SMIF protein, a key p...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(02)02751-5

    authors: Callebaut I

    更新日期:2002-05-22 00:00:00

  • Complete amino acid sequence of human seminal plasma beta-inhibin. Prediction of post Gln-Arg cleavage as a maturation site.

    abstract::The complete sequence of a 94 amino acid human seminal plasma polypeptide exhibiting inhibin-like activity is presented. This molecule, called beta-inhibin, selectively and specifically suppresses the release of pituitary FSH in vivo as well as in vitro. It does not affect the secretion of LH. Such a novel acidic prot...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(84)80766-8

    authors: Seidah NG,Arbatti NJ,Rochemont J,Sheth AR,Chrétien M

    更新日期:1984-10-01 00:00:00

  • NTR1 encodes a high affinity oligopeptide transporter in Arabidopsis.

    abstract::Hterologous complementation of yeast mutants has enabled the isolation of genes encoding several families of amino acid transporters. Among them, NTR1 codes for a membrane protein with weak histidine transport activity. However, at the sequence level, NTR1 is related to rather non-specific oligopeptide transporters fr...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(95)00853-2

    authors: Rentsch D,Laloi M,Rouhara I,Schmelzer E,Delrot S,Frommer WB

    更新日期:1995-08-21 00:00:00

  • Both plant and animal LEA proteins act as kinetic stabilisers of polyglutamine-dependent protein aggregation.

    abstract::LEA (late embryogenesis abundant) proteins are intrinsically disordered proteins that contribute to stress tolerance in plants and invertebrates. Here we show that, when both plant and animal LEA proteins are co-expressed in mammalian cells with self-aggregating polyglutamine (polyQ) proteins, they reduce aggregation ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2011.01.020

    authors: Liu Y,Chakrabortee S,Li R,Zheng Y,Tunnacliffe A

    更新日期:2011-02-18 00:00:00

  • Requirements for a conservative protein translocation pathway in chloroplasts.

    abstract::The chloroplast inner envelope translocon subunit Tic110 is imported via a soluble stromal translocation intermediate. In this study an in-organellar import system is established which allows for an accumulation of this intermediate in order to analyze its requirements for reexport. All results demonstrate that the re...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2007.05.004

    authors: Vojta L,Soll J,Bölter B

    更新日期:2007-06-12 00:00:00

  • Gene duplication in bovine brain myelin proteolipid and homology with related proteins.

    abstract::Analysis of the amino acid sequence of bovine brain myelin proteolipid reveals not only extensive internal homology, but also homology with portions of the myelin basic protein, the peripheral nerve myelin protein, Po, and with the small proteolipid subunit of mitochondrial ATP synthase. These results suggest that the...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(83)80732-7

    authors: Laursen RA,Samiullah M,Lees MB

    更新日期:1983-09-05 00:00:00

  • BIT, an immune antigen receptor-like molecule in the brain.

    abstract::We previously found a brain-specific glycoprotein in the rat brain. It postnatally increases and is rich in the mature brain. We cloned cDNA of this protein. It is composed of a signal peptide, a V-type immunoglobulin domain, two C1-type immunoglobulin domains, a transmembrane segment and a cytoplasmic region containi...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(97)00724-2

    authors: Sano S,Ohnishi H,Omori A,Hasegawa J,Kubota M

    更新日期:1997-07-14 00:00:00

  • Highly selective affinity labeling of the primer-binding site of E. coli DNA polymerase I.

    abstract::Highly selective affinity labeling of the primer site of E. coli DNA polymerase I was performed with the 5'-reactive derivatives of oligothymidylate in the presence of poly(dA) template. Subtilysine cleavage proved that the site of affinity modification belonged to the 'Klenow' part of DNA polymerase I. If taken separ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(90)80478-2

    authors: Mitina RL,Mustaev AA,Zaychikov EF,Khomov VV,Lavrik OI

    更新日期:1990-10-15 00:00:00

  • Inhibition of Na+-dependent Ca2+ efflux from heart mitochondria by amiloride analogues.

    abstract::The Na+-induced release of accumulated Ca2+ from heart mitochondria is inhibited by amiloride, benzamil and several other amiloride analogues. These drugs do not affect uptake or release of Ca2+ mediated by the ruthenium red-sensitive uniporter and their effects, like those of diltiazem and other Ca2+-antagonists, app...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(83)80768-6

    authors: Jurkowitz MS,Altschuld RA,Brierley GP,Cragoe EJ Jr

    更新日期:1983-10-17 00:00:00

  • Electrons generated by photosystem II are utilized by an oxidase in the absence of photosystem I in the cyanobacterium Synechocystis sp. PCC 6803.

    abstract::The reduction and reoxidation kinetics of the first quinone-type electron acceptor in photosystem II, QA-, were measured by fluorescence in a light-tolerant, photosystem I-less strain of the cyanobacterium Synechocystis sp. PCC 6803. In this strain, which shows excellent amplitudes of variable fluorescence, the rate o...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(94)80638-1

    authors: Vermaas WF,Shen G,Styring S

    更新日期:1994-01-03 00:00:00

  • Inositol(1,3,4,5) tetrakisphosphate plays an important role in calcium mobilization from Entamoeba histolytica.

    abstract::Calcium release from internal stores of Entamoeba histolytica, a parasitic protozoan, was observed by measuring fluorescence of Fura-2. Emptying of inositol(1,4,5)trisphosphate (Ins(1,4,5)P3)-sensitive calcium pools in permeabilized E. histolytica did not significantly affect subsequent calcium release by inositol(1,3...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(95)00265-b

    authors: Raha S,Giri B,Bhattacharyya B,Biswas BB

    更新日期:1995-04-10 00:00:00

  • Crystal structure of human nuclear pore complex component NUP43.

    abstract::Nuclear pore complexes (NPC) form nuclear pores that cross the nuclear envelope and allow molecules to transport between the nucleus and the cytoplasm. We solved the crystal structure of human Nup43 (hNUP43), an important component in the Nup107 subcomplex of NPC. hNup43 adopts a seven-bladed β-propeller fold. We conf...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2015.09.008

    authors: Xu C,Li Z,He H,Wernimont A,Li Y,Loppnau P,Min J

    更新日期:2015-10-24 00:00:00

  • Production of alkenes and novel secondary products by P450 OleTJE using novel H2 O2 -generating fusion protein systems.

    abstract::Jeotgalicoccus sp. 8456 OleTJE (CYP152L1) is a fatty acid decarboxylase cytochrome P450 that uses hydrogen peroxide (H2 O2 ) to catalyse production of terminal alkenes, which are industrially important chemicals with biofuel applications. We report enzyme fusion systems in which Streptomyces coelicolor alditol oxidase...

    journal_title:FEBS letters

    pub_type: 信件

    doi:10.1002/1873-3468.12581

    authors: Matthews S,Tee KL,Rattray NJ,McLean KJ,Leys D,Parker DA,Blankley RT,Munro AW

    更新日期:2017-03-01 00:00:00

  • Human release factor eRF1: structural organisation of the unique functional gene on chromosome 5 and of the three processed pseudogenes.

    abstract::In lower and higher eukaryotes, a family of tightly related proteins designated eRF1 (for eukaryotic release factor 1) catalyses termination of protein synthesis at all three stop codons. The human genome contains four eRF1 homologous sequences localised on chromosomes 5, 6, 7 and X. We report here the cloning and the...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(99)00795-4

    authors: Guenet L,Toutain B,Guilleret I,Chauvel B,Deaven LL,Longmire JL,Le Gall JY,David V,Le Treut A

    更新日期:1999-07-02 00:00:00

  • Preparation and structure of the charge-transfer intermediate of the transmembrane redox catalyst DsbB.

    abstract::Disulfide bond formation is a critical step in the folding of many secretory proteins. In bacteria, disulfide bonds are introduced by the periplasmic dithiol oxidase DsbA, which transfers its catalytic disulfide bond to folding polypeptides. Reduced DsbA is reoxidized by ubiquinone Q8, catalyzed by inner membrane quin...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2008.07.063

    authors: Malojcić G,Owen RL,Grimshaw JP,Glockshuber R

    更新日期:2008-10-15 00:00:00

  • Positive feedback between ethanolamine-specific phospholipid base exchange and cytochrome P450 activities in rat liver microsomes. The effect of clofibric acid.

    abstract::The results of the present investigation relate the effects of the nutritional state and administration of clofibric acid (CLA), a hypolipidaemic drug and peroxisomal proliferator, on phosphatidylethanolamine (PE) synthesis in rat liver and fatty acid metabolism. Fasting and CLA treatment of animals causes an increase...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(98)00960-0

    authors: Lenart J,Komańska I,Pikuła S,Jasińska R

    更新日期:1998-08-28 00:00:00

  • YFH1-mediated iron homeostasis is independent of mitochondrial respiration.

    abstract::The human gene frataxin and its yeast homolog YFH1 affect mitochondrial function. Deficits in frataxin result in Friedreich ataxia, while deletion of YFH1 results in respiratory incompetence. We determined that as long as respiratory incompetent yeast express Yfh1p they do not accumulate excessive mitochondrial iron. ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(01)03137-4

    authors: Chen OS,Kaplan J

    更新日期:2001-11-30 00:00:00

  • Mass spectrometry studies of demetallation of haemin by recombinant horse L chain apoferritin and its mutant (E 53,56,57,60 Q).

    abstract::An essential difference between eukaryotic ferritins and bacterioferritins is that the latter contain naturally, in vivo haem as Fe-protoporphyrin IX. This haem is located in a hydrophobic pocket along the 2-fold symmetry axes and is liganded by two Met 52. However, in in vivo studies, a cofactor has been isolated in ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2006.10.034

    authors: de Val N,Herschbach H,Potier N,Dorsselaer AV,Crichton RR

    更新日期:2006-11-13 00:00:00

  • Mutations in subunit 6 of the F1F0-ATP synthase cause two entirely different diseases.

    abstract::A lowered efficiency of oxidative phosphorylation was recently found in a Leber hereditary optic neuropathy (LHON) proband carrying a mutation in the mtDNA gene for subunit 6 of the membrane-bound F0 segment of the F1F0-ATP synthase [9]. This phenotype was transferred to cytoplasmic hybrid cells together with the muta...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(97)00757-6

    authors: Majander A,Lamminen T,Juvonen V,Aula P,Nikoskelainen E,Savontaus ML,Wikström M

    更新日期:1997-07-28 00:00:00

  • Functional site of bukatoxin, an alpha-type sodium channel neurotoxin from the Chinese scorpion (Buthus martensi Karsch) venom: probable role of the (52)PDKVP(56) loop.

    abstract::Alpha-toxins from scorpion venoms prolong the action potential of excitable cells by blocking sodium channel inactivation. We have purified bukatoxin, an alpha-toxin from scorpion (Buthus martensi Karsch) venom, to homogeneity. Bukatoxin produced marked relaxant responses in the carbachol-precontracted rat anococcygeu...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(01)02342-0

    authors: Srinivasan KN,Nirthanan S,Sasaki T,Sato K,Cheng B,Gwee MC,Kini RM,Gopalakrishnakone P

    更新日期:2001-04-13 00:00:00

  • Activation of leukocyte rolling by the cysteine-rich domain and the hyper-variable region of HF3, a snake venom hemorrhagic metalloproteinase.

    abstract::The functionality of the disintegrin-like/cysteine-rich domains of snake venom metalloproteinases (SVMPs) has been shown to reside in the cysteine-rich region, which can interact with VWA-containing proteins. Recently, the hyper-variable region (HVR) of the cysteine-rich domain was suggested to constitute a potential ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2008.10.034

    authors: Menezes MC,Paes Leme AF,Melo RL,Silva CA,Della Casa M,Bruni FM,Lima C,Lopes-Ferreira M,Camargo AC,Fox JW,Serrano SM

    更新日期:2008-11-26 00:00:00

  • Mitochondrial functional interactions between frataxin and Isu1p, the iron-sulfur cluster scaffold protein, in Saccharomyces cerevisiae.

    abstract::Friedreich's ataxia is caused by a deficit in the mitochondrial protein frataxin. The present work demonstrates that in vivo yeast frataxin Yfh1p and Isu1p, the mitochondrial scaffold protein for the Fe-S cluster assembly, have tightly linked biological functions, acting in concert to promote the Fe-S cluster assembly...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(03)01498-4

    authors: Ramazzotti A,Vanmansart V,Foury F

    更新日期:2004-01-16 00:00:00

  • In vivo modulation of rat liver acyl-coenzyme A:cholesterol acyltransferase by phosphorylation and substrate supply.

    abstract::Rats were infused intravenously with a bolus of mevalonolactone or cholesterol-rich chylomicron and microsomes prepared from the livers at times up to 2 h after the infusion. Acyl-CoA cholesterol acyltransferase (ACAT) activity was measured in the microsomes under conditions that define the activity state of the enzym...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(83)80410-4

    authors: Suckling KE,Stange EF,Dietschy JM

    更新日期:1983-08-08 00:00:00