Cloning of thioredoxin h reductase and characterization of the thioredoxin reductase-thioredoxin h system from wheat
Thioredoxins h are ubiquitous proteins reduced by NADPH- thioredoxin reductase (NTR). They are able to reduce disulphides in target proteins. In monocots, thioredoxins h accumulate at high level in seeds and show a predominant localization in the nucleus of seed cells. These results suggest that the...
Saved in:
Published in | Biochemical journal Vol. 367; no. Pt 2; pp. 491 - 497 |
---|---|
Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
England
15.10.2002
|
Subjects | |
Online Access | Get full text |
Cover
Loading…
Abstract | Thioredoxins h are ubiquitous proteins reduced by NADPH- thioredoxin reductase (NTR). They are able to reduce disulphides in target proteins. In monocots, thioredoxins h accumulate at high level in seeds and show a predominant localization in the nucleus of seed cells. These results suggest that the NTR-thioredoxin h system probably plays an important role in seed physiology. To date, the study of this system in monocots is limited by the lack of information about NTR. In the present study, we describe the cloning of a full-length cDNA encoding NTR from wheat ( Triticum aestivum ). The polypeptide deduced from this cDNA shows close similarity to NTRs from Arabidopsis, contains FAD- and NADPH-binding domains and a disulphide probably interacting with the disulphide at the active site of thioredoxin h. Wheat NTR was expressed in Escherichia coli as a His-tagged protein. The absorption spectrum of the purified recombinant protein is typical of flavoenzymes. Furthermore, it showed NADPH-dependent thioredoxin h reduction activity, thus confirming that the cDNA clone reported in the present study encodes wheat NTR. Using the His-tagged NTR and TRXhA (wheat thioredoxin h ), we successfully reconstituted the wheat NTR-thioredoxin h system in vitro, as shown by the insulin reduction assay. A polyclonal antibody was raised against wheat NTR after immunization of rabbits with the purified His-tagged protein. This antibody efficiently detected a single polypeptide of the corresponding molecular mass in seed extracts and it allowed the analysis of the pattern of accumulation of NTR in different wheat organs and developmental stages. NTR shows a wide distribution in wheat, but, surprisingly, its accumulation in seeds is low, in contrast with the level of thioredoxins h. |
---|---|
AbstractList | Thioredoxins h are ubiquitous proteins reduced by NADPH- thioredoxin reductase (NTR). They are able to reduce disulphides in target proteins. In monocots, thioredoxins h accumulate at high level in seeds and show a predominant localization in the nucleus of seed cells. These results suggest that the NTR-thioredoxin h system probably plays an important role in seed physiology. To date, the study of this system in monocots is limited by the lack of information about NTR. In the present study, we describe the cloning of a full-length cDNA encoding NTR from wheat ( Triticum aestivum ). The polypeptide deduced from this cDNA shows close similarity to NTRs from Arabidopsis, contains FAD- and NADPH-binding domains and a disulphide probably interacting with the disulphide at the active site of thioredoxin h. Wheat NTR was expressed in Escherichia coli as a His-tagged protein. The absorption spectrum of the purified recombinant protein is typical of flavoenzymes. Furthermore, it showed NADPH-dependent thioredoxin h reduction activity, thus confirming that the cDNA clone reported in the present study encodes wheat NTR. Using the His-tagged NTR and TRXhA (wheat thioredoxin h ), we successfully reconstituted the wheat NTR-thioredoxin h system in vitro, as shown by the insulin reduction assay. A polyclonal antibody was raised against wheat NTR after immunization of rabbits with the purified His-tagged protein. This antibody efficiently detected a single polypeptide of the corresponding molecular mass in seed extracts and it allowed the analysis of the pattern of accumulation of NTR in different wheat organs and developmental stages. NTR shows a wide distribution in wheat, but, surprisingly, its accumulation in seeds is low, in contrast with the level of thioredoxins h. |
Author | Serrato, Antonio J Pérez-Ruiz, Juan M Cejudo, Francisco J |
AuthorAffiliation | Instituto de Bioquímica Vegetal y Fotosíntesis, Centro de Investigaciones Científicas Isla de la Cartuja, Avda Américo Vespucio s/n, 41092 Seville, Spain |
AuthorAffiliation_xml | – name: Instituto de Bioquímica Vegetal y Fotosíntesis, Centro de Investigaciones Científicas Isla de la Cartuja, Avda Américo Vespucio s/n, 41092 Seville, Spain |
Author_xml | – sequence: 1 givenname: Antonio J surname: Serrato fullname: Serrato, Antonio J organization: Instituto de Bioquímica Vegetal y Fotosíntesis, Centro de Investigaciones Científicas Isla de la Cartuja, Avda Américo Vespucio s/n, 41092 Seville, Spain – sequence: 2 givenname: Juan M surname: Pérez-Ruiz fullname: Pérez-Ruiz, Juan M – sequence: 3 givenname: Francisco J surname: Cejudo fullname: Cejudo, Francisco J |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/12106017$$D View this record in MEDLINE/PubMed |
BookMark | eNpVkUtLxDAQx4Mouj4ufgDpyYNQnTy2aS-CLL5A8KLnkKYTG2kTTbK-Pr0ru6h7moH5zW8G_rtk0wePhBxSOKUg2Fn7zAAYUOAbZEKFhLKWrN4kE2CVKCtgdIfspvQMQAUI2CY7lFGogMoJybMheOefimCL3LsQsQsfzhd9sejmJuuEhfZdYXodtckY3ZfOLvglj2s7vxvluil9poxjYWMYi_cedd4nW1YPCQ9WdY88Xl0-zG7Ku_vr29nFXWmEgFwanKLlMG2ngja8aVrRGWYtIrct55WQtQTbWTSsxppLU7e1lgKN7jQV067le-R86X2ZtyN2Bn2OelAv0Y06fqqgnVqfeNerp_CmKGOsbuRCcLwSxPA6x5TV6JLBYdAewzwpyaioGuAL8GQJmhhSimh_j1BQPyGpv5AW8NH_t_7QVSr8G5Omkx0 |
CitedBy_id | crossref_primary_10_3389_fpls_2019_00380 crossref_primary_10_1016_j_fob_2015_10_001 crossref_primary_10_1016_j_ijbiomac_2014_12_033 crossref_primary_10_1016_j_plaphy_2004_03_002 crossref_primary_10_1074_jbc_M704298200 crossref_primary_10_1007_s11032_016_0563_y crossref_primary_10_1016_j_plantsci_2015_12_002 crossref_primary_10_1016_j_biochi_2004_10_008 crossref_primary_10_1104_pp_110_171082 crossref_primary_10_1139_g04_057 crossref_primary_10_1016_j_jplph_2012_08_014 crossref_primary_10_1093_jxb_ern037 crossref_primary_10_1093_mp_ssp025 crossref_primary_10_32615_bp_2020_155 crossref_primary_10_1007_s00253_016_7688_0 crossref_primary_10_1104_pp_107_113639 crossref_primary_10_1104_pp_110_170712 crossref_primary_10_1016_j_plantsci_2017_02_001 crossref_primary_10_1016_j_crvi_2007_01_001 crossref_primary_10_1016_j_jcs_2009_04_002 crossref_primary_10_1016_j_plaphy_2011_06_009 crossref_primary_10_1104_pp_111_177196 crossref_primary_10_1016_j_fgb_2023_103799 crossref_primary_10_1016_j_bbagen_2012_08_020 crossref_primary_10_1016_j_plantsci_2015_03_016 crossref_primary_10_1093_mp_ssn096 crossref_primary_10_1016_j_bbapap_2016_02_014 crossref_primary_10_1016_j_febslet_2014_09_044 crossref_primary_10_3389_fpls_2016_00883 crossref_primary_10_1186_1471_2229_14_110 crossref_primary_10_1021_jf303513m |
ContentType | Journal Article |
DBID | CGR CUY CVF ECM EIF NPM AAYXX CITATION 7X8 5PM |
DOI | 10.1042/bj20020103 |
DatabaseName | Medline MEDLINE MEDLINE (Ovid) MEDLINE MEDLINE PubMed CrossRef MEDLINE - Academic PubMed Central (Full Participant titles) |
DatabaseTitle | MEDLINE Medline Complete MEDLINE with Full Text PubMed MEDLINE (Ovid) CrossRef MEDLINE - Academic |
DatabaseTitleList | MEDLINE - Academic CrossRef MEDLINE |
Database_xml | – sequence: 1 dbid: NPM name: PubMed url: https://proxy.k.utb.cz/login?url=http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed sourceTypes: Index Database – sequence: 2 dbid: EIF name: MEDLINE url: https://proxy.k.utb.cz/login?url=https://www.webofscience.com/wos/medline/basic-search sourceTypes: Index Database |
DeliveryMethod | fulltext_linktorsrc |
Discipline | Anatomy & Physiology Chemistry |
EISSN | 1470-8728 |
EndPage | 497 |
ExternalDocumentID | 10_1042_bj20020103 12106017 |
Genre | Research Support, Non-U.S. Gov't Journal Article |
GroupedDBID | --- -DZ -~X .55 .GJ 0R~ 23N 2WC 3EH 3O- 4.4 53G 5GY 5RE 5VS 6J9 79B AABGO AAHRG ABPPZ ABTAH ACGFO ACGFS ACNCT ADBBV AEGXH AENEX AI. AIAGR AIZAD ALMA_UNASSIGNED_HOLDINGS BAWUL CGR CS3 CUY CVF DU5 E3Z EBS ECM EIF EJD F20 F5P H13 HH6 HYE HZ~ K-O L7B MV1 MVM N9A NPM O9- OHT OK1 P2P RHI RNS RPM RPO TR2 TWZ VH1 WH7 WOQ X7M XOL XSW Y6R YNY ZGI ZXP ZY4 ~02 ~KM AAYXX CITATION 7X8 5PM |
ID | FETCH-LOGICAL-c440t-ce5ef305b5419399b4dc2ffee3fb33647870fdfec28e837c8b8a74ecada145db3 |
IEDL.DBID | RPM |
ISSN | 0264-6021 |
IngestDate | Tue Sep 17 20:53:45 EDT 2024 Thu Aug 15 23:02:21 EDT 2024 Fri Aug 23 01:25:26 EDT 2024 Sat Sep 28 07:37:07 EDT 2024 |
IsDoiOpenAccess | false |
IsOpenAccess | true |
IsPeerReviewed | true |
IsScholarly | true |
Issue | Pt 2 |
Language | English |
LinkModel | DirectLink |
MergedId | FETCHMERGED-LOGICAL-c440t-ce5ef305b5419399b4dc2ffee3fb33647870fdfec28e837c8b8a74ecada145db3 |
Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
OpenAccessLink | https://europepmc.org/articles/pmc1222897?pdf=render |
PMID | 12106017 |
PQID | 72146903 |
PQPubID | 23479 |
PageCount | 7 |
ParticipantIDs | pubmedcentral_primary_oai_pubmedcentral_nih_gov_1222897 proquest_miscellaneous_72146903 crossref_primary_10_1042_bj20020103 pubmed_primary_12106017 |
PublicationCentury | 2000 |
PublicationDate | 2002-Oct-15 2002-10-15 20021015 |
PublicationDateYYYYMMDD | 2002-10-15 |
PublicationDate_xml | – month: 10 year: 2002 text: 2002-Oct-15 day: 15 |
PublicationDecade | 2000 |
PublicationPlace | England |
PublicationPlace_xml | – name: England |
PublicationTitle | Biochemical journal |
PublicationTitleAlternate | Biochem J |
PublicationYear | 2002 |
References | 11012662 - Eur J Biochem. 2000 Oct;267(20):6110-7 2449095 - Anal Biochem. 1987 Nov 1;166(2):368-79 9108027 - Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3621-6 11488482 - Plant Mol Biol. 2001 Jun;46(3):361-71 8308900 - J Mol Biol. 1994 Jan 28;235(4):1357-63 15012197 - Annu Rev Plant Physiol Plant Mol Biol. 2000 Jun;51:371-400 9006939 - J Biol Chem. 1997 Jan 31;272(5):2936-41 9108029 - Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3633-8 11536727 - Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3169-75 10497208 - J Biol Chem. 1999 Oct 1;274(40):28459-65 8577704 - Proc Natl Acad Sci U S A. 1996 Feb 6;93(3):1006-11 10344197 - Plant Mol Biol. 1999 Mar;39(5):915-26 10947986 - Science. 2000 Aug 18;289(5482):1190-4 7777559 - Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5620-4 8650234 - Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):6146-51 9501259 - Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):3312-7 11717467 - Proc Natl Acad Sci U S A. 2001 Nov 20;98(24):14144-9 7476354 - Methods Enzymol. 1995;252:199-208 1910303 - Arch Biochem Biophys. 1991 Jul;288(1):1-9 2067578 - Nature. 1991 Jul 11;352(6331):172-4 10037727 - J Biol Chem. 1999 Mar 5;274(10):6366-73 9523703 - Eur J Biochem. 1998 Mar 1;252(2):314-24 9000629 - J Mol Biol. 1996 Dec 20;264(5):1044-57 10215850 - Eur J Biochem. 1999 Apr;261(2):405-12 |
References_xml | |
SSID | ssj0014040 |
Score | 1.9682024 |
Snippet | Thioredoxins h are ubiquitous proteins reduced by NADPH- thioredoxin reductase (NTR). They are able to reduce disulphides in target proteins. In monocots,... Thioredoxins h are ubiquitous proteins reduced by NADPH— thioredoxin reductase (NTR). They are able to reduce disulphides in target proteins. In monocots,... |
SourceID | pubmedcentral proquest crossref pubmed |
SourceType | Open Access Repository Aggregation Database Index Database |
StartPage | 491 |
SubjectTerms | Amino Acid Sequence Animals Cloning, Molecular DNA, Complementary Escherichia coli - genetics Gene Expression Regulation, Plant Insulin - metabolism Kinetics Molecular Sequence Data Phylogeny Rabbits Recombinant Proteins - genetics Recombinant Proteins - metabolism Sequence Homology, Amino Acid Thioredoxin h Thioredoxin-Disulfide Reductase - genetics Thioredoxin-Disulfide Reductase - immunology Thioredoxin-Disulfide Reductase - metabolism Thioredoxins - genetics Thioredoxins - metabolism Triticum - genetics |
Title | Cloning of thioredoxin h reductase and characterization of the thioredoxin reductase-thioredoxin h system from wheat |
URI | https://www.ncbi.nlm.nih.gov/pubmed/12106017 https://search.proquest.com/docview/72146903 https://pubmed.ncbi.nlm.nih.gov/PMC1222897 |
Volume | 367 |
hasFullText | 1 |
inHoldings | 1 |
isFullTextHit | |
isPrint | |
link | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwnV1JTwIxFH5BLnoxCi64NtF4K8zSmWGOSCSEBONBEm6Taec1YKQQHKL-e9tZ0NGbt0m6ZNL32rd_D-BWmB7RPiK1vdiiTEhBQ0cg7doutyWToeWb2uHxoz-csNHUm9bAK2thsqR9wedt9bpoq_ksy61cLUSnzBPrPI37tnFbhEFnB3YC1y1N9CJ0wCxWOFYY9bUEKzFJmdO5H5mUBNPZwGCFamNHWyNBVSD90TJ_J0v-kD6DA9gv1EbSy3_vEGqoGtDsKW0yLz7JHckSOTMPeQN2-2UTtyak_dfM30qWkqSz-dLAg37MFZmRtcFsTbUMI7FKiNjiNudlmfl8rKzZrqDVnXIwaGIKVci7eduPYDJ4eO4PadFogQrGrJQK9FDqi889pvW5MOQsEY6UiK7krgGY15daJhKF00Vt0Iou78YBQxEnsc28hLvHUFdLhadATEiY-_phEDxh3Pd47CLqz1gbRq5IvBbclKcdrXI8jSiLgzMn4i8leVpwXRIi0gdmYhixwuXmLQpMI_LQzDjJyfK9S0HPFgQVgm0nGCDt6ojmrwxQu-Cns3-vPIe9vEuMpXn-AurpeoOXWllJ-VXGnF8bIO2x |
link.rule.ids | 230,315,733,786,790,891,27957,27958,53827,53829 |
linkProvider | National Library of Medicine |
linkToHtml | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwnV1LT9wwEB7xOMCFFih0-wBLRb15Nw8n2Rzpqmh5LOoBELcodsbapawXQVZ9_PqOk3hp4AS3SPZYScb2vL8BOFC2R3SMyP0o97hQWvE0UMj7fih9LXTqxbZ2eHQeDy_FyXV0vQSRq4WpkvaVnHTN7bRrJuMqt_JuqnouT6z3YzTwrdsiTXrLsErnNUickd4ED4QnGteK4DHJMIdKKoLetxOblGB7G1i0UDJ3yB5J2iLpmZ75NF3yP_lz9Aau3JvXaSc_u_NSdtXfJ6COL_60t7DRaKTssB7ehCU0W7B9aMgan_5hX1mVI1o537dgbeD6w21DObitXLlsplk5nsws8ujviWFjdm_hYEsSjyw3BVMLSOi64rOejy2aBQVvr1TjTDNbA8N-WbHxDi6Pvl8Mhrzp4cCVEF7JFUao6U6RkSBVMU2lKFSgNWKoZWix6-m-0IVGFfSRbGXVl_08EajyIvdFVMhwB1bMzOB7YDbaLGO6c5QshIwjmYeI9JiTzRWqIurAF8fG7K6G6siqELsIMnnj-N6BfcfhjH6YDY_kBmfzhyyxPc5TO2O35vfjKs1G6UDS2gmLCRajuz1C_K2wuht-fng15T6sDS9GZ9nZ8fnpR1ivm9F4dLQ-wUp5P8fPpBOVcq86Af8ASVUPtw |
linkToPdf | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwnV1LT9wwEB5RkFouhUILy9MSVW_evJxkc4SFFaWAOBQJ9RLFzli7hfWuaFYt_HrGeWwJ3LhFyjhKMrbn9fkbgK_K9oiOELkXZi4XSiue-Ap5zwukp4VO3MieHb64jE6vxdlNePOs1VcJ2ldy1DV3464ZDUts5XSsnAYn5lxd9D2btkhiZ5pr5x0s0Zr1kyZQrwsIwhV1ekXwiOxYw0wqfOfozAITbH8DyxhKIQ_FJHHbLL3yNV9CJp_ZoMEK_GrevoKe3HZnheyqxxfEjm_6vFX4WHum7LAS-QQLaNZg_dBQVD5-YN9YiRUtk_Br8KHf9Ilbh6J_V6Z02USzYjiaWAbSfyPDhuze0sIWZCZZZnKm5tTQ1cnPSh5bY-YjePtJFd80s2dh2F9rPj7D9eDkZ_-U170cuBLCLbjCEDXtLTIU5DImiRS58rVGDLQMLIc97Rs616j8HlLMrHqyl8UCVZZnnghzGXyBRTMxuAnMVp1lRHuPkrmQUSizAJEuM4q9ApWHHThoVJlOK8qOtCy1Cz-Vvxvdd2C_0XJKP8yWSTKDk9mfNLa9zhMrsVHp_P9T6snSgbg1G-YClqu7fYd0XHJ21zrdevPIfXh_dTxIz79f_tiG5aonjUsrbAcWi_sZ7pJrVMi9chE8Aat5Ejc |
openUrl | ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=Cloning+of+thioredoxin+h+reductase+and+characterization+of+the+thioredoxin+reductase-thioredoxin+h+system+from+wheat&rft.jtitle=Biochemical+journal&rft.au=Serrato%2C+Antonio+J&rft.au=P%C3%A9rez-Ruiz%2C+Juan+M&rft.au=Cejudo%2C+Francisco+J&rft.date=2002-10-15&rft.issn=0264-6021&rft.eissn=1470-8728&rft.volume=367&rft.issue=Pt+2&rft.spage=491&rft.epage=497&rft_id=info:doi/10.1042%2FBJ20020103&rft_id=info%3Apmid%2F12106017&rft.externalDBID=PMC1222897 |
thumbnail_l | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/lc.gif&issn=0264-6021&client=summon |
thumbnail_m | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/mc.gif&issn=0264-6021&client=summon |
thumbnail_s | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/sc.gif&issn=0264-6021&client=summon |