OrangerV1, PascalFrancis, Curation, bibRecord, 000483

Gene isolation, analysis of expression, and in vitro synthesis of glutathione S-transferase from orange fruit [Citrus sinensis L. (Osbeck)]

Identifieur interne : 000483 ( PascalFrancis/Curation ); précédent : 000482; suivant : 000484

Gene isolation, analysis of expression, and in vitro synthesis of glutathione S-transferase from orange fruit [Citrus sinensis L. (Osbeck)]

Auteurs : Angela Roberta Lo Piero [Italie] ; Ivana Puglisi [Italie] ; Goffredo Petrone [Italie]

Source :

Journal of agricultural and food chemistry : (Print) [ 0021-8561 ] ; 2006.

RBID : Pascal:07-0005154

Descripteurs français

Pascal (Inist)
- Expression génique, Isolement, In vitro, Citrus sinensis, Synthèse, Glutathione transferase, Orange, Agrume, Flavonoïde, Anthocyane, Sang, Froid, Stress, Temps réel, Réaction chaîne polymérase.
Wicri :
- topic : Agrume.

English descriptors

KwdEn :
- Anthocyanin, Blood, Citrus fruit, Citrus sinensis, Cold, Flavonoid, Gene expression, Glutathione transferase, In vitro, Isolation, Orange, Polymerase chain reaction, Real time, Stress, Synthesis.

Abstract

Glutathione S-transferases (GSTs) (EC 2.5.1.18) are ubiquitous enzymes that have a defined role in xenobiotic detoxification, but a deeper knowledge of their function in endogenous metabolism is still lacking. In this work, we isolated the cDNAs as well as the genomic clones of orange GSTs. Having considered gene organization and homology data, we suggest that the isolated GSTgene is probably involved in the vacuolar import of anthocyanins. We also found that the blood and blond orange GSTs shared the same nucleotide sequences, but as expected, the GST expression in the nonpigmented orange cultivar [Citrus sinensis L. (Osbeck)] (Navel and Ovale) was strongly reduced as compared to that of the pigmented orange (Tarocco). Interestingly, in the crude extracts of pigmented orange fruits, the GSTactivity was reproducibly detected by providing either 1-chloro-2,4 dinitrobenzene (CDNB) or cyanidin-3-O-glucoside (C-3-G) as substrates; moreover, we have shown that cyanidin-3-O-glucoside acted as a powerful competitive inhibitor of 1-chloro-2,4 dinitrobenzene conjugation to reduced glutathione (GSH) in the pigmented orange, confirming that this molecule might easily bind to the active site of the enzyme and functions as a putative substrate. In addition, we have reported here the successful in vitro expression of orange GSTcDNAs leading to a GST enzyme that is active against cyanidin-3-O-glucoside, thus suggesting the probable involvement of the isolated gene in the tagging of anthocyanins for vacuolar import. This last result will help to study the kinetic and structural properties of orange fruit GSTavoiding time-consuming protein purification procedures.

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A03		`1`		`@0 J. agric. food chem. : (Print)`
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A08	`01`	`1`	`ENG`	`@1 Gene isolation, analysis of expression, and in vitro synthesis of glutathione S-transferase from orange fruit [Citrus sinensis L. (Osbeck)]`
A11	`01`	`1`		`@1 LO PIERO (Angela Roberta)`
A11	`02`	`1`		`@1 PUGLISI (Ivana)`
A11	`03`	`1`		`@1 PETRONE (Goffredo)`
A14	`01`			`@1 Dipartimento di Scienze Agronomiche, Agrochimiche e delle Produzioni Animali (DACPA), Facoltà di Agraria, Università degli Studi Catania, via S. Sofia no. 98 @2 95123 Catania @3 ITA @Z 1 aut. @Z 2 aut. @Z 3 aut.`
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C01	`01`		`ENG`	@0 Glutathione S-transferases (GSTs) (EC 2.5.1.18) are ubiquitous enzymes that have a defined role in xenobiotic detoxification, but a deeper knowledge of their function in endogenous metabolism is still lacking. In this work, we isolated the cDNAs as well as the genomic clones of orange GSTs. Having considered gene organization and homology data, we suggest that the isolated GSTgene is probably involved in the vacuolar import of anthocyanins. We also found that the blood and blond orange GSTs shared the same nucleotide sequences, but as expected, the GST expression in the nonpigmented orange cultivar [Citrus sinensis L. (Osbeck)] (Navel and Ovale) was strongly reduced as compared to that of the pigmented orange (Tarocco). Interestingly, in the crude extracts of pigmented orange fruits, the GSTactivity was reproducibly detected by providing either 1-chloro-2,4 dinitrobenzene (CDNB) or cyanidin-3-O-glucoside (C-3-G) as substrates; moreover, we have shown that cyanidin-3-O-glucoside acted as a powerful competitive inhibitor of 1-chloro-2,4 dinitrobenzene conjugation to reduced glutathione (GSH) in the pigmented orange, confirming that this molecule might easily bind to the active site of the enzyme and functions as a putative substrate. In addition, we have reported here the successful in vitro expression of orange GSTcDNAs leading to a GST enzyme that is active against cyanidin-3-O-glucoside, thus suggesting the probable involvement of the isolated gene in the tagging of anthocyanins for vacuolar import. This last result will help to study the kinetic and structural properties of orange fruit GSTavoiding time-consuming protein purification procedures.
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C03	`01`	`X`	`ENG`	`@0 Gene expression @5 01`
C03	`01`	`X`	`SPA`	`@0 Expresión genética @5 01`
C03	`02`	`X`	`FRE`	`@0 Isolement @5 02`
C03	`02`	`X`	`ENG`	`@0 Isolation @5 02`
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C03	`03`	`X`	`ENG`	`@0 In vitro @5 10`
C03	`03`	`X`	`SPA`	`@0 In vitro @5 10`
C03	`04`	`X`	`FRE`	`@0 Citrus sinensis @2 NS @5 13`
C03	`04`	`X`	`ENG`	`@0 Citrus sinensis @2 NS @5 13`
C03	`04`	`X`	`SPA`	`@0 Citrus sinensis @2 NS @5 13`
C03	`05`	`X`	`FRE`	`@0 Synthèse @5 19`
C03	`05`	`X`	`ENG`	`@0 Synthesis @5 19`
C03	`05`	`X`	`SPA`	`@0 Síntesis @5 19`
C03	`06`	`X`	`FRE`	`@0 Glutathione transferase @2 FE @5 20`
C03	`06`	`X`	`ENG`	`@0 Glutathione transferase @2 FE @5 20`
C03	`06`	`X`	`SPA`	`@0 Glutathione transferase @2 FE @5 20`
C03	`07`	`X`	`FRE`	`@0 Orange @5 24`
C03	`07`	`X`	`ENG`	`@0 Orange @5 24`
C03	`07`	`X`	`SPA`	`@0 Naranja @5 24`
C03	`08`	`X`	`FRE`	`@0 Agrume @5 26`
C03	`08`	`X`	`ENG`	`@0 Citrus fruit @5 26`
C03	`08`	`X`	`SPA`	`@0 Agrios @5 26`
C03	`09`	`X`	`FRE`	`@0 Flavonoïde @5 28`
C03	`09`	`X`	`ENG`	`@0 Flavonoid @5 28`
C03	`09`	`X`	`SPA`	`@0 Flavonoide @5 28`
C03	`10`	`X`	`FRE`	`@0 Anthocyane @5 48`
C03	`10`	`X`	`ENG`	`@0 Anthocyanin @5 48`
C03	`10`	`X`	`SPA`	`@0 Antocianina @5 48`
C03	`11`	`X`	`FRE`	`@0 Sang @5 53`
C03	`11`	`X`	`ENG`	`@0 Blood @5 53`
C03	`11`	`X`	`SPA`	`@0 Sangre @5 53`
C03	`12`	`X`	`FRE`	`@0 Froid @5 63`
C03	`12`	`X`	`ENG`	`@0 Cold @5 63`
C03	`12`	`X`	`SPA`	`@0 Frío @5 63`
C03	`13`	`X`	`FRE`	`@0 Stress @5 64`
C03	`13`	`X`	`ENG`	`@0 Stress @5 64`
C03	`13`	`X`	`SPA`	`@0 Estrés @5 64`
C03	`14`	`X`	`FRE`	`@0 Temps réel @5 65`
C03	`14`	`X`	`ENG`	`@0 Real time @5 65`
C03	`14`	`X`	`SPA`	`@0 Tiempo real @5 65`
C03	`15`	`X`	`FRE`	`@0 Réaction chaîne polymérase @5 66`
C03	`15`	`X`	`ENG`	`@0 Polymerase chain reaction @5 66`
C03	`15`	`X`	`SPA`	`@0 Reacción cadena polimerasa @5 66`
C07	`01`	`X`	`FRE`	`@0 Fruit @5 08`
C07	`01`	`X`	`ENG`	`@0 Fruit @5 08`
C07	`01`	`X`	`SPA`	`@0 Fruto @5 08`
C07	`02`	`X`	`FRE`	`@0 Rutaceae @2 NS`
C07	`02`	`X`	`ENG`	`@0 Rutaceae @2 NS`
C07	`02`	`X`	`SPA`	`@0 Rutaceae @2 NS`
C07	`03`	`X`	`FRE`	`@0 Dicotyledones @2 NS`
C07	`03`	`X`	`ENG`	`@0 Dicotyledones @2 NS`
C07	`03`	`X`	`SPA`	`@0 Dicotyledones @2 NS`
C07	`04`	`X`	`FRE`	`@0 Angiospermae @2 NS`
C07	`04`	`X`	`ENG`	`@0 Angiospermae @2 NS`
C07	`04`	`X`	`SPA`	`@0 Angiospermae @2 NS`
C07	`05`	`X`	`FRE`	`@0 Spermatophyta @2 NS`
C07	`05`	`X`	`ENG`	`@0 Spermatophyta @2 NS`
C07	`05`	`X`	`SPA`	`@0 Spermatophyta @2 NS`
C07	`06`	`X`	`FRE`	`@0 Transferases @2 FE`
C07	`06`	`X`	`ENG`	`@0 Transferases @2 FE`
C07	`06`	`X`	`SPA`	`@0 Transferases @2 FE`
C07	`07`	`X`	`FRE`	`@0 Enzyme @2 FE`
C07	`07`	`X`	`ENG`	`@0 Enzyme @2 FE`
C07	`07`	`X`	`SPA`	`@0 Enzima @2 FE`
C07	`08`	`X`	`FRE`	`@0 Pigment @5 49`
C07	`08`	`X`	`ENG`	`@0 Pigments @5 49`
C07	`08`	`X`	`SPA`	`@0 Pigmento @5 49`
C07	`09`	`X`	`FRE`	`@0 Facteur milieu @5 50`
C07	`09`	`X`	`ENG`	`@0 Environmental factor @5 50`
C07	`09`	`X`	`SPA`	`@0 Factor medio @5 50`
C07	`10`	`X`	`FRE`	`@0 Température @5 51`
C07	`10`	`X`	`ENG`	`@0 Temperature @5 51`
C07	`10`	`X`	`SPA`	`@0 Temperatura @5 51`
C07	`11`	`X`	`FRE`	`@0 Biologie moléculaire @5 52`
C07	`11`	`X`	`ENG`	`@0 Molecular biology @5 52`
C07	`11`	`X`	`SPA`	`@0 Biología molecular @5 52`
C07	`12`	`X`	`FRE`	`@0 Polyphénol @5 75`
C07	`12`	`X`	`ENG`	`@0 Polyphenol @5 75`
C07	`12`	`X`	`SPA`	`@0 Polifenol @5 75`
N21				`@1 008`
N44	`01`			`@1 OTO`
N82				`@1 OTO`

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Le document en format XML

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<front><div type="abstract" xml:lang="en">Glutathione S-transferases (GSTs) (EC 2.5.1.18) are ubiquitous enzymes that have a defined role in xenobiotic detoxification, but a deeper knowledge of their function in endogenous metabolism is still lacking. In this work, we isolated the cDNAs as well as the genomic clones of orange GSTs. Having considered gene organization and homology data, we suggest that the isolated GSTgene is probably involved in the vacuolar import of anthocyanins. We also found that the blood and blond orange GSTs shared the same nucleotide sequences, but as expected, the GST expression in the nonpigmented orange cultivar [Citrus sinensis L. (Osbeck)] (Navel and Ovale) was strongly reduced as compared to that of the pigmented orange (Tarocco). Interestingly, in the crude extracts of pigmented orange fruits, the GSTactivity was reproducibly detected by providing either 1-chloro-2,4 dinitrobenzene (CDNB) or cyanidin-3-O-glucoside (C-3-G) as substrates; moreover, we have shown that cyanidin-3-O-glucoside acted as a powerful competitive inhibitor of 1-chloro-2,4 dinitrobenzene conjugation to reduced glutathione (GSH) in the pigmented orange, confirming that this molecule might easily bind to the active site of the enzyme and functions as a putative substrate. In addition, we have reported here the successful in vitro expression of orange GSTcDNAs leading to a GST enzyme that is active against cyanidin-3-O-glucoside, thus suggesting the probable involvement of the isolated gene in the tagging of anthocyanins for vacuolar import. This last result will help to study the kinetic and structural properties of orange fruit GSTavoiding time-consuming protein purification procedures.</div>
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<s5>01</s5>
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<fC03 i1="02" i2="X" l="FRE"><s0>Isolement</s0>
<s5>02</s5>
</fC03>
<fC03 i1="02" i2="X" l="ENG"><s0>Isolation</s0>
<s5>02</s5>
</fC03>
<fC03 i1="02" i2="X" l="SPA"><s0>Aislamiento</s0>
<s5>02</s5>
</fC03>
<fC03 i1="03" i2="X" l="FRE"><s0>In vitro</s0>
<s5>10</s5>
</fC03>
<fC03 i1="03" i2="X" l="ENG"><s0>In vitro</s0>
<s5>10</s5>
</fC03>
<fC03 i1="03" i2="X" l="SPA"><s0>In vitro</s0>
<s5>10</s5>
</fC03>
<fC03 i1="04" i2="X" l="FRE"><s0>Citrus sinensis</s0>
<s2>NS</s2>
<s5>13</s5>
</fC03>
<fC03 i1="04" i2="X" l="ENG"><s0>Citrus sinensis</s0>
<s2>NS</s2>
<s5>13</s5>
</fC03>
<fC03 i1="04" i2="X" l="SPA"><s0>Citrus sinensis</s0>
<s2>NS</s2>
<s5>13</s5>
</fC03>
<fC03 i1="05" i2="X" l="FRE"><s0>Synthèse</s0>
<s5>19</s5>
</fC03>
<fC03 i1="05" i2="X" l="ENG"><s0>Synthesis</s0>
<s5>19</s5>
</fC03>
<fC03 i1="05" i2="X" l="SPA"><s0>Síntesis</s0>
<s5>19</s5>
</fC03>
<fC03 i1="06" i2="X" l="FRE"><s0>Glutathione transferase</s0>
<s2>FE</s2>
<s5>20</s5>
</fC03>
<fC03 i1="06" i2="X" l="ENG"><s0>Glutathione transferase</s0>
<s2>FE</s2>
<s5>20</s5>
</fC03>
<fC03 i1="06" i2="X" l="SPA"><s0>Glutathione transferase</s0>
<s2>FE</s2>
<s5>20</s5>
</fC03>
<fC03 i1="07" i2="X" l="FRE"><s0>Orange</s0>
<s5>24</s5>
</fC03>
<fC03 i1="07" i2="X" l="ENG"><s0>Orange</s0>
<s5>24</s5>
</fC03>
<fC03 i1="07" i2="X" l="SPA"><s0>Naranja</s0>
<s5>24</s5>
</fC03>
<fC03 i1="08" i2="X" l="FRE"><s0>Agrume</s0>
<s5>26</s5>
</fC03>
<fC03 i1="08" i2="X" l="ENG"><s0>Citrus fruit</s0>
<s5>26</s5>
</fC03>
<fC03 i1="08" i2="X" l="SPA"><s0>Agrios</s0>
<s5>26</s5>
</fC03>
<fC03 i1="09" i2="X" l="FRE"><s0>Flavonoïde</s0>
<s5>28</s5>
</fC03>
<fC03 i1="09" i2="X" l="ENG"><s0>Flavonoid</s0>
<s5>28</s5>
</fC03>
<fC03 i1="09" i2="X" l="SPA"><s0>Flavonoide</s0>
<s5>28</s5>
</fC03>
<fC03 i1="10" i2="X" l="FRE"><s0>Anthocyane</s0>
<s5>48</s5>
</fC03>
<fC03 i1="10" i2="X" l="ENG"><s0>Anthocyanin</s0>
<s5>48</s5>
</fC03>
<fC03 i1="10" i2="X" l="SPA"><s0>Antocianina</s0>
<s5>48</s5>
</fC03>
<fC03 i1="11" i2="X" l="FRE"><s0>Sang</s0>
<s5>53</s5>
</fC03>
<fC03 i1="11" i2="X" l="ENG"><s0>Blood</s0>
<s5>53</s5>
</fC03>
<fC03 i1="11" i2="X" l="SPA"><s0>Sangre</s0>
<s5>53</s5>
</fC03>
<fC03 i1="12" i2="X" l="FRE"><s0>Froid</s0>
<s5>63</s5>
</fC03>
<fC03 i1="12" i2="X" l="ENG"><s0>Cold</s0>
<s5>63</s5>
</fC03>
<fC03 i1="12" i2="X" l="SPA"><s0>Frío</s0>
<s5>63</s5>
</fC03>
<fC03 i1="13" i2="X" l="FRE"><s0>Stress</s0>
<s5>64</s5>
</fC03>
<fC03 i1="13" i2="X" l="ENG"><s0>Stress</s0>
<s5>64</s5>
</fC03>
<fC03 i1="13" i2="X" l="SPA"><s0>Estrés</s0>
<s5>64</s5>
</fC03>
<fC03 i1="14" i2="X" l="FRE"><s0>Temps réel</s0>
<s5>65</s5>
</fC03>
<fC03 i1="14" i2="X" l="ENG"><s0>Real time</s0>
<s5>65</s5>
</fC03>
<fC03 i1="14" i2="X" l="SPA"><s0>Tiempo real</s0>
<s5>65</s5>
</fC03>
<fC03 i1="15" i2="X" l="FRE"><s0>Réaction chaîne polymérase</s0>
<s5>66</s5>
</fC03>
<fC03 i1="15" i2="X" l="ENG"><s0>Polymerase chain reaction</s0>
<s5>66</s5>
</fC03>
<fC03 i1="15" i2="X" l="SPA"><s0>Reacción cadena polimerasa</s0>
<s5>66</s5>
</fC03>
<fC07 i1="01" i2="X" l="FRE"><s0>Fruit</s0>
<s5>08</s5>
</fC07>
<fC07 i1="01" i2="X" l="ENG"><s0>Fruit</s0>
<s5>08</s5>
</fC07>
<fC07 i1="01" i2="X" l="SPA"><s0>Fruto</s0>
<s5>08</s5>
</fC07>
<fC07 i1="02" i2="X" l="FRE"><s0>Rutaceae</s0>
<s2>NS</s2>
</fC07>
<fC07 i1="02" i2="X" l="ENG"><s0>Rutaceae</s0>
<s2>NS</s2>
</fC07>
<fC07 i1="02" i2="X" l="SPA"><s0>Rutaceae</s0>
<s2>NS</s2>
</fC07>
<fC07 i1="03" i2="X" l="FRE"><s0>Dicotyledones</s0>
<s2>NS</s2>
</fC07>
<fC07 i1="03" i2="X" l="ENG"><s0>Dicotyledones</s0>
<s2>NS</s2>
</fC07>
<fC07 i1="03" i2="X" l="SPA"><s0>Dicotyledones</s0>
<s2>NS</s2>
</fC07>
<fC07 i1="04" i2="X" l="FRE"><s0>Angiospermae</s0>
<s2>NS</s2>
</fC07>
<fC07 i1="04" i2="X" l="ENG"><s0>Angiospermae</s0>
<s2>NS</s2>
</fC07>
<fC07 i1="04" i2="X" l="SPA"><s0>Angiospermae</s0>
<s2>NS</s2>
</fC07>
<fC07 i1="05" i2="X" l="FRE"><s0>Spermatophyta</s0>
<s2>NS</s2>
</fC07>
<fC07 i1="05" i2="X" l="ENG"><s0>Spermatophyta</s0>
<s2>NS</s2>
</fC07>
<fC07 i1="05" i2="X" l="SPA"><s0>Spermatophyta</s0>
<s2>NS</s2>
</fC07>
<fC07 i1="06" i2="X" l="FRE"><s0>Transferases</s0>
<s2>FE</s2>
</fC07>
<fC07 i1="06" i2="X" l="ENG"><s0>Transferases</s0>
<s2>FE</s2>
</fC07>
<fC07 i1="06" i2="X" l="SPA"><s0>Transferases</s0>
<s2>FE</s2>
</fC07>
<fC07 i1="07" i2="X" l="FRE"><s0>Enzyme</s0>
<s2>FE</s2>
</fC07>
<fC07 i1="07" i2="X" l="ENG"><s0>Enzyme</s0>
<s2>FE</s2>
</fC07>
<fC07 i1="07" i2="X" l="SPA"><s0>Enzima</s0>
<s2>FE</s2>
</fC07>
<fC07 i1="08" i2="X" l="FRE"><s0>Pigment</s0>
<s5>49</s5>
</fC07>
<fC07 i1="08" i2="X" l="ENG"><s0>Pigments</s0>
<s5>49</s5>
</fC07>
<fC07 i1="08" i2="X" l="SPA"><s0>Pigmento</s0>
<s5>49</s5>
</fC07>
<fC07 i1="09" i2="X" l="FRE"><s0>Facteur milieu</s0>
<s5>50</s5>
</fC07>
<fC07 i1="09" i2="X" l="ENG"><s0>Environmental factor</s0>
<s5>50</s5>
</fC07>
<fC07 i1="09" i2="X" l="SPA"><s0>Factor medio</s0>
<s5>50</s5>
</fC07>
<fC07 i1="10" i2="X" l="FRE"><s0>Température</s0>
<s5>51</s5>
</fC07>
<fC07 i1="10" i2="X" l="ENG"><s0>Temperature</s0>
<s5>51</s5>
</fC07>
<fC07 i1="10" i2="X" l="SPA"><s0>Temperatura</s0>
<s5>51</s5>
</fC07>
<fC07 i1="11" i2="X" l="FRE"><s0>Biologie moléculaire</s0>
<s5>52</s5>
</fC07>
<fC07 i1="11" i2="X" l="ENG"><s0>Molecular biology</s0>
<s5>52</s5>
</fC07>
<fC07 i1="11" i2="X" l="SPA"><s0>Biología molecular</s0>
<s5>52</s5>
</fC07>
<fC07 i1="12" i2="X" l="FRE"><s0>Polyphénol</s0>
<s5>75</s5>
</fC07>
<fC07 i1="12" i2="X" l="ENG"><s0>Polyphenol</s0>
<s5>75</s5>
</fC07>
<fC07 i1="12" i2="X" l="SPA"><s0>Polifenol</s0>
<s5>75</s5>
</fC07>
<fN21><s1>008</s1>
</fN21>
<fN44 i1="01"><s1>OTO</s1>
</fN44>
<fN82><s1>OTO</s1>
</fN82>
</pA>
</standard>
</inist>
</record>

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   |texte=   Gene isolation, analysis of expression, and in vitro synthesis of glutathione S-transferase from orange fruit [Citrus sinensis L. (Osbeck)]
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Gene isolation, analysis of expression, and in vitro synthesis of glutathione S-transferase from orange fruit [Citrus sinensis L. (Osbeck)]

Gene isolation, analysis of expression, and in vitro synthesis of glutathione S-transferase from orange fruit [Citrus sinensis L. (Osbeck)]

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