Identification of the molecular basis for the functional difference between flavonoid 3'-hydroxylase and flavonoid 3',5'-hydroxylase.


:Flavonoid 3'-hydroxylase (F3'H) and flavonoid 3',5'-hydroxylase (F3'5'H) are cytochrome P450 enzymes and determine the B-ring hydroxylation pattern of flavonoids by introducing hydroxyl groups at the 3'- or the 3'- and 5'-position, respectively. Sequence identity between F3'H and F3'5'H is generally low since their divergence took place early in the evolution of higher plants. However, in the Asteraceae the family-specific evolution of an F3'5'H from an F3'H precursor occurred, and consequently sequence identity is substantially higher. We used this phenomenon for alignment studies, in order to identify regions which could be involved in determining substrate specificity and functionality. Subsequent construction and expression of chimeric genes indicated that substrate specificity of F3'H and F3'5'H is determined near the N-terminal end and the functional difference between these two enzymes near the C-terminal end. The impact on function of individual amino acids located in substrate recognition site 6 (SRS6) was further tested by site-directed mutagenesis. Most interestingly, a conservative Thr to Ser exchange at position 487 conferred additional 5'-hydroxylation activity to recombinant Gerbera hybrida F3'H, whereas the reverse substitution transformed recombinant Osteospermum hybrida F3'5'H into an F3'H with low remaining 5'-hydroxylation activity. Since the physicochemical properties of Thr and Ser are highly similar, the difference in size appears to be the main factor contributing to functional difference. The results further suggest that relatively few amino acids exchanges were required for the evolutionary extension of 3'- to 3',5'-hydroxylation activity.




FEBS letters


Seitz C,Ameres S,Forkmann G




Has Abstract


2007-07-24 00:00:00














  • Activation of boar proacrosin is effected by processing at both N- and C-terminal portions of the zymogen molecule.

    abstract::A mixture of 55 and 53 kDa boar proacrosins was autoactivated at pH 8.5 to produce a 43 kDa intermediate form and a 35 kDa mature acrosin, and each of four forms of (pro)acrosins was isolated. Analysis of the N-terminal sequences of the two proacrosins indicated the existence of a segment corresponding to the acrosin ...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Baba T,Michikawa Y,Kawakura K,Arai Y

    更新日期:1989-02-13 00:00:00

  • A large conductance ion channel in the nuclear envelope of a higher plant cell.

    abstract::To detect and characterize ion channel activity in the nuclear envelope of a higher plant cell, we performed patch clamp experiments on nuclei isolated from coconut endosperm cells and on giant liposomes containing nuclear envelope fragments prepared from the same cells. An ion channel exhibiting a number of conductan...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Matzke AJ,Behensky C,Weiger T,Matzke MA

    更新日期:1992-05-04 00:00:00

  • Six myosin heavy chain isoforms are expressed during chick breast muscle development.

    abstract::Two major embryonic myosin heavy chains are expressed in embryonic chick breast muscle until the first week after hatching. Of these, one is already detected in the 8-day-old embryo. The other appears on day 12. Two putative slow embryonic isoforms represent minor components transiently expressed between days 8 and 12...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Hofmann S,Düsterhöft S,Pette D

    更新日期:1988-10-10 00:00:00

  • One of the three proteinase inhibitor genes newly identified in the Brassica napus genome codes for an inhibitor of glutamyl endopeptidase.

    abstract::Three proteinase inhibitor genes have been identified in the rapeseed (Brassica napus) genome. They are highly homologous to other genes of the mustard inhibitor (MSI) family of proteinase inhibitors characteristic of Cruciferae. In germinating seeds, only the transcript of one gene, coding for a trypsin inhibitor, is...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: De Leo F,Volpicella M,Sciancalepore M,Gallerani R,Ceci LR

    更新日期:2006-02-06 00:00:00

  • A leucine-rich repeat peptide derived from the Drosophila Toll receptor forms extended filaments with a beta-sheet structure.

    abstract::Leucine-rich repeats (LRRs) are 22-28 amino acid-long sequence motifs found in a family of cytoplasmic, membrane and extracellular proteins. There is evidence that LRRs function in signal transduction, cellular adhesion and protein-protein interactions. Here we report unusual properties of a synthetic LRR peptide deri...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Gay NJ,Packman LC,Weldon MA,Barna JC

    更新日期:1991-10-07 00:00:00

  • Platelet-activating factor mobilizes intracellular calcium in vascular smooth muscle cells.

    abstract::The effect of platelet-activating factor (PAF) on polyphosphoinositide metabolism and 45Ca2+ efflux was examined in a vascular smooth muscle cell line (A7r5). PAF stimulated a rapid but transient production of inositol trisphosphate and inositol bisphosphate which, in the presence of lithium, resulted in an accumulati...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Doyle VM,Creba JA,Rüegg UT

    更新日期:1986-03-03 00:00:00

  • The interaction of fructose 2,6-bisphosphate with an allosteric site of rat liver fructose 1,6-bisphosphatase.

    abstract::Rat liver fructose 1,6-bisphosphatase can be protected against partial inactivation by N-ethylmaleimide by low concentrations of fructose 2,6-bisphosphate or high concentrations of fructose 1,6-bisphosphate. The partially inactivated enzyme has a much reduced sensitivity to high substrate inhibition and has lost the s...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Meek DW,Nimmo HG

    更新日期:1983-08-22 00:00:00

  • Calcium signaling mediates the response to cadmium toxicity in Saccharomyces cerevisiae cells.

    abstract::The involvement of Ca(2+) in the response to high Mn(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+), Cd(2+), and Hg(2+) was investigated in Saccharomyces cerevisiae. The yeast cells responded through a sharp increase in cytosolic Ca(2+) when exposed to Cd(2+), and to a lesser extent to Cu(2+), but not to Mn(2+), Co(2+), Ni(2+), ...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Ruta LL,Popa VC,Nicolau I,Danet AF,Iordache V,Neagoe AD,Farcasanu IC

    更新日期:2014-08-25 00:00:00

  • The orientations of core antenna chlorophylls in photosystem II are optimized to maximize the quantum yield of photosynthesis.

    abstract::In photosystem II (PSII) the probability that energy absorbed by core antenna chlorophyll (Chl) is transferred to the reaction center (RC) is extremely high. Although close proximity between antenna Chl ensures a high transfer efficiency, relative pigment orientation can fractionally modify it. This level of refinemen...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Vasil'ev S,Shen JR,Kamiya N,Bruce D

    更新日期:2004-03-12 00:00:00

  • Human REG family genes are tandemly ordered in a 95-kilobase region of chromosome 2p12.

    abstract::Reg, first isolated from a rat regenerating islet cDNA library, is expressed in regenerating islet beta-cells. Recently, it has been revealed that Reg and Reg-related genes constitute a multigene family, the Reg family. In human, the four REG family genes, i.e., REG 1 alpha, REG 1 beta, REG-related sequence (RS) and H...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Miyashita H,Nakagawara K,Mori M,Narushima Y,Noguchi N,Moriizumi S,Takasawa S,Yonekura H,Takeuchi T,Okamoto H

    更新日期:1995-12-27 00:00:00

  • Subcellular localization of the pyoverdine biogenesis machinery of Pseudomonas aeruginosa: a membrane-associated "siderosome".

    abstract::The peptidic siderophore pyoverdine is the primary iron uptake system of fluorescent pseudomonads, and a virulence factor in the opportunistic pathogen Pseudomonas aeruginosa. Pyoverdine biogenesis is a co-ordinate process requiring several precursor-generating enzymes and large nonribosomal peptide synthetases (NRPSs...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Imperi F,Visca P

    更新日期:2013-11-01 00:00:00

  • Alternative splicing of the human Shaker K+ channel beta 1 gene and functional expression of the beta 2 gene product.

    abstract::Mammalian voltage-activated Shaker K+ channels associate with at least three cytoplasmic proteins: Kv beta 1, Kv beta 2 and Kv beta 3. These beta subunits contain variable N-termini, which can modulate the inactivation of Shaker alpha subunits, but are homologous throughout an aldo-keto reductase core. Human and ferre...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: McCormack K,McCormack T,Tanouye M,Rudy B,Stühmer W

    更新日期:1995-08-14 00:00:00

  • Reduced maximum capacity of glycolysis in brown adipose tissue of genetically obese, diabetic (db/db) mice and its restoration following treatment with a thermogenic beta-adrenoceptor agonist.

    abstract::The maximal activities of the key glycolytic enzymes hexokinase and 6-phosphofructokinase, were reduced in brown adipose tissue in db/db mice compared to their lean littermates. Treatment of db/db mice with the thermogenic beta-adrenoceptor agonist, BRL 26830, restored normoglycaemia. The only significant increase in ...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Young P,Cawthorne MA,Levy AL,Wilson K

    更新日期:1984-10-15 00:00:00

  • The N-terminal region of the alpha-subunit of the TRAP complex has a conserved cluster of negative charges.

    abstract::The alpha-subunit of the TRAP complex (TRAP alpha) is a single-spanning membrane protein of the endoplasmic reticulum (ER) which is found in proximity of nascent polypeptide chains translocating across the membrane. Here, we demonstrate the widespread occurrence of TRAP alpha in eukaryotes as indicated by its existenc...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Hartmann E,Prehn S

    更新日期:1994-08-08 00:00:00

  • Delineation of the TRAK binding regions of the kinesin-1 motor proteins.

    abstract::Understanding specific cargo distribution in differentiated cells is a major challenge. Trafficking kinesin proteins (TRAKs) are kinesin adaptors. They bind the cargo binding domain of kinesin-1 motor proteins forming a link between the motor and their cargoes. To refine the TRAK1/2 binding sites within the kinesin-1 ...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Randall TS,Moores C,Stephenson FA

    更新日期:2013-11-29 00:00:00

  • Urmylation and tRNA thiolation functions of ubiquitin-like Uba4·Urm1 systems are conserved from yeast to man.

    abstract::The ubiquitin-like protein Urm1 from budding yeast and its E1-like activator Uba4 have dual roles in protein urmylation and tRNA thiolation pathways. To study whether these are conserved among eukaryotes, we used gene shuffles to replace the yeast proteins by their human counterparts, hURM1 and hUBA4/MOCS3. As judged ...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Jüdes A,Ebert F,Bär C,Thüring KL,Harrer A,Klassen R,Helm M,Stark MJ,Schaffrath R

    更新日期:2015-04-02 00:00:00

  • Vapor-phase modification of sulfhydryl groups in proteins.

    abstract::Proteins and peptides are readily and specifically modified at their sulfhydryl groups by the vapors of a mixture of 4-vinylpyridine and tributylphosphine. The phenylthiohydantoin derivative of S-beta-(4-pyridylethyl)cysteine formed during sequence analysis is easily detectable in current identification systems. ...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Amons R

    更新日期:1987-02-09 00:00:00

  • Human manganese superoxide dismutase suppresses HER2/neu-mediated breast cancer malignancy.

    abstract::The up-regulation of HER2/neu is associated with human malignancies and is a useful target for developing anticancer drugs. Overexpression of human manganese superoxide dismutase (MnSOD) has been demonstrated to effectively suppress various carcinoma cells, including breast carcinomas, in vitro and in vivo. This study...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Chuang TC,Liu JY,Lin CT,Tang YT,Yeh MH,Chang SC,Li JW,Kao MC

    更新日期:2007-09-18 00:00:00

  • Human CyP33 binds specifically to mRNA and binding stimulates PPIase activity of hCyP33.

    abstract::Human nuclear cyclophilin 33 (hCyP33) was the first protein which was found to contain an RNA-binding motif and a PPIase domain. It was not known what cellular and physiological roles are played by the RNA-binding activity as well as the PPIase activity of hCyP33. In this paper, we investigated the binding specificity...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Wang Y,Han R,Zhang W,Yuan Y,Zhang X,Long Y,Mi H

    更新日期:2008-03-05 00:00:00

  • Activation of Na,K-ATPase by an endogenous peptide, PEC-60.

    abstract::Several peptidic and non-peptidic factors can modulate Na,K-ATPase activity, among them mainly inhibitors of this enzyme, ouabain being the most effective. In a very few cases only, activation of Na,K-ATPase by endogenous factors has been recorded. We have investigated the effect on Na,K-ATPase of a novel regulatory p...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Kairane C,Zilmer M,Mutt V,Sillard R

    更新日期:1994-05-23 00:00:00

  • Isolation and analysis of the mouse opsin gene.

    abstract::We have identified three overlapping 5'-truncated mouse opsin cDNA clones by immunologically screening a lambda gt11 retina expression library. Using one of the cDNA clones as a probe, we isolated a 5 kb genomic fragment that encompassed the complete coding sequence for mouse opsin. The coding region for opsin was int...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Baehr W,Falk JD,Bugra K,Triantafyllos JT,McGinnis JF

    更新日期:1988-10-10 00:00:00

  • Inhibition of cellular response to platelet-derived growth factor by low M(r) phosphotyrosine protein phosphatase overexpression.

    abstract::The role of low M(r) phosphotyrosine protein phosphatase (PTPase) in the control of cell proliferation was studied. A synthetic gene coding for PTPase was transfected and expressed in NIH/3T3 fibroblasts. The effects of the enzyme were particularly evident when cells were stimulated by platelet-derived growth factor (...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Berti A,Rigacci S,Raugei G,Degl'Innocenti D,Ramponi G

    更新日期:1994-07-25 00:00:00

  • Rat glutathione transferase 8-8, an enzyme efficiently detoxifying 4-hydroxyalk-2-enals.

    abstract::Rat glutathione transferase 8-8 is one of the less abundant cytosolic glutathione transferases, accounting for approx. 1% of the total activity with 1-chloro-2,4-dinitrobenzene in liver. The enzyme is eluted at pH 6.3 upon chromatofocusing and has so far been identified in liver, kidney, lung and testis. Characteristi...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Jensson H,Guthenberg C,Alin P,Mannervik B

    更新日期:1986-07-28 00:00:00

  • Acylation of adenylyl cyclase catalyst is important for enzymic activity.

    abstract::Incubation of human thrombocytes in the presence of [3H]palmitic acid leads to incorporation of this fatty acid into the alpha subunit of Gs as described [Linder et al., Proc. Natl. Acad. Sci. USA 90 (1993) 3675-3679; Degtyarev et al., Biochemistry 32 (1993) 8057-8061] but also into the catalyst of adenylyl cyclase wh...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Mollner S,Beck K,Pfeuffer T

    更新日期:1995-09-11 00:00:00

  • Specific inhibition of Na-Ca exchange function by antisense oligodeoxynucleotides.

    abstract::The Na-Ca exchanger is essential for the Ca2+ homeostasis in many cell types. This transporter has been difficult to investigate because no specific inhibitor is available. We have synthesized an antisense oligodeoxynucleotide directed against the rat cardiac Na-Ca exchanger mRNA. To estimate the activity of the Na-Ca...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Lipp P,Schwaller B,Niggli E

    更新日期:1995-05-08 00:00:00

  • Identification of the amino acid residues involved in selective agonist binding in the first extracellular loop of the delta- and mu-opioid receptors.

    abstract::Effects of amino acid substitutions in the first extracellular loop region of the delta- and mu-opioid receptors were examined. Substitution of lysine-108 of the delta-receptor (delta K108) with asparagine improved affinity to [D-Ala2,MePhe4,Gly-ol5]enk ephalin (DAGO), a mu-selective peptide agonist, to be comparable ...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Fukuda K,Terasako K,Kato S,Mori K

    更新日期:1995-10-09 00:00:00

  • A new approach for EPR detection of hydroxyl radicals by reaction with sterically hindered cyclic amines and oxygen.

    abstract::Sterically hindered cyclic amines react with hydroxyl radicals in the presence of oxygen to yield stable nitroxide radicals which can be detected by EPR. This reaction provides a nonconventional spin-trapping tool for detection of hydroxyl radicals. ...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Rosenthal I,Krishna CM,Yang GC,Kondo T,Riesz P

    更新日期:1987-09-28 00:00:00

  • A specific immunological probe for the major myelin proteolipid. Confirmation of a deletion in DM-20.

    abstract::Major myelin proteolipid (MMPL, also called PLP) and DM-20 are the two major intrinsic membrane proteins of CNS myelin. A specific immunological probe was obtained for MMPL by raising antibodies against the synthetic tridecapeptide 117-129 of MMPL. Antibodies against this peptide reacted with the MMPL but did not cros...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Trifilieff E,Luu B,Nussbaum JL,Roussel G,Espinosa de los Monteros A,Sabatier JM,Van Rietschoten J

    更新日期:1986-03-31 00:00:00

  • The Methanocaldococcus jannaschii protein Mj0968 is not a P-type ATPase.

    abstract::The Methanocaldococcus jannaschii (formerly Methanococcus jannaschii) protein Mj0968 has been reported to represent a soluble P-type ATPase [Ogawa et al., FEBS Lett. 471 (2000) 99-102]. In this study, we report that the heterologously expressed Mj0968-His(10) protein exhibits high rates of phosphatase activity, wherea...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Bramkamp M,Gassel M,Herkenhoff-Hesselmann B,Bertrand J,Altendorf K

    更新日期:2003-05-22 00:00:00

  • Biphasic effect of fructose 2,6-bisphosphate on the liver fructose-1,6-bisphosphatase: mechanistic and physiological implications.

    abstract::Fructose 2,6-bisphosphate has been claimed to be both a substrate analogue and an allosteric inhibitor of fructose-1,6-bisphosphatase. The results reported here show that fructose 2,6-bisphosphate can be both an inhibitor and an activator of the enzyme, depending on the substrate concentration. This biphasic behaviour...

    journal_title:FEBS letters

    pub_type: 杂志文章


    authors: Corredor C,Boscá L,Sols A

    更新日期:1984-02-27 00:00:00