Analysis of the wild-type and mutant genes encoding the enzyme kynurenine monooxygenase of the yellow fever mosquito, Aedes aegypti
Kynurenine 3-monooxygenase (KMO) catalyses the hydroxylation of kynurenine to 3-hydroxykynurenine. KMO has a key role in tryptophan catabolism and synthesis of ommochrome pigments in mosquitoes. The gene encoding this enzyme in the yellow fever mosquito, Aedes aegypti, is called kynurenine hydroxyla...
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| Veröffentlicht in: | Insect molecular biology 2003-10, Vol.12 (5), p.483-490 |
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| creator | Han, Q Calvo, E Marinotti, O Fang, J Rizzi, M James, A.A Li, J |
| description | Kynurenine 3-monooxygenase (KMO) catalyses the hydroxylation of kynurenine to 3-hydroxykynurenine. KMO has a key role in tryptophan catabolism and synthesis of ommochrome pigments in mosquitoes. The gene encoding this enzyme in the yellow fever mosquito, Aedes aegypti, is called kynurenine hydroxylase (kh) and a mutant allele that produces white eyes has been designated khw. A number of cDNA clones representative of wild-type and mutant genes were isolated. Sequence analyses of the wild-type and mutant cDNAs revealed a deletion of 162 nucleotides in the mutant gene near the 3'-end of the deduced coding region. RT-PCR analyses confirm the transcription of a truncated mRNA in the mutant strain. The in-frame deletion results in a loss of 54 amino acids, which disrupts a major α-helix and which probably accounts for the loss of activity of the enzyme. Recombinant Ae. aegypti KMO showed high substrate specificity for kynurenine with optimum activity at 40 °C and pH = 7.5. Kinetic parameters and inhibition of KMO activity by Cl- and pyridoxal-5-phosphate were determined. |
| doi_str_mv | 10.1046/j.1365-2583.2003.00433.x |
| format | Article |
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KMO has a key role in tryptophan catabolism and synthesis of ommochrome pigments in mosquitoes. The gene encoding this enzyme in the yellow fever mosquito, Aedes aegypti, is called kynurenine hydroxylase (kh) and a mutant allele that produces white eyes has been designated khw. A number of cDNA clones representative of wild-type and mutant genes were isolated. Sequence analyses of the wild-type and mutant cDNAs revealed a deletion of 162 nucleotides in the mutant gene near the 3'-end of the deduced coding region. RT-PCR analyses confirm the transcription of a truncated mRNA in the mutant strain. The in-frame deletion results in a loss of 54 amino acids, which disrupts a major α-helix and which probably accounts for the loss of activity of the enzyme. Recombinant Ae. aegypti KMO showed high substrate specificity for kynurenine with optimum activity at 40 °C and pH = 7.5. Kinetic parameters and inhibition of KMO activity by Cl- and pyridoxal-5-phosphate were determined.</description><identifier>ISSN: 0962-1075</identifier><identifier>EISSN: 1365-2583</identifier><identifier>DOI: 10.1046/j.1365-2583.2003.00433.x</identifier><identifier>PMID: 12974953</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Science Ltd</publisher><subject>3-hydroxykynurenine ; Aedes - enzymology ; Aedes - genetics ; Aedes aegypti ; amino acid sequences ; Animals ; complementary DNA ; Culicidae ; deletion mutants ; DNA, Complementary - genetics ; DNA, Complementary - isolation & purification ; Electrophoresis, Polyacrylamide Gel ; enzyme activity ; enzyme inhibition ; enzyme kinetics ; Gene Deletion ; gene expression ; Gene Expression Profiling ; insect vectors ; Kinetics ; KMO gene ; Kynurenine ; Kynurenine 3-Monooxygenase ; kynurenine hydroxylase ; messenger RNA ; Mixed Function Oxygenases - chemistry ; Mixed Function Oxygenases - genetics ; molecular sequence data ; mutants ; mutation ; nucleotide sequences ; open reading frames ; oxygenases ; recombinant proteins ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment ; structural genes ; substrate specificity ; truncation mutants ; white eye ; Yellow fever virus</subject><ispartof>Insect molecular biology, 2003-10, Vol.12 (5), p.483-490</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c6283-d7c7586f00924eb02241e8c3f3a1121e60cdd9689a7f1ec43eb3caded36ba40a3</citedby><cites>FETCH-LOGICAL-c6283-d7c7586f00924eb02241e8c3f3a1121e60cdd9689a7f1ec43eb3caded36ba40a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1046%2Fj.1365-2583.2003.00433.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1046%2Fj.1365-2583.2003.00433.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>230,314,776,780,881,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12974953$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Han, Q</creatorcontrib><creatorcontrib>Calvo, E</creatorcontrib><creatorcontrib>Marinotti, O</creatorcontrib><creatorcontrib>Fang, J</creatorcontrib><creatorcontrib>Rizzi, M</creatorcontrib><creatorcontrib>James, A.A</creatorcontrib><creatorcontrib>Li, J</creatorcontrib><title>Analysis of the wild-type and mutant genes encoding the enzyme kynurenine monooxygenase of the yellow fever mosquito, Aedes aegypti</title><title>Insect molecular biology</title><addtitle>Insect Mol Biol</addtitle><description>Kynurenine 3-monooxygenase (KMO) catalyses the hydroxylation of kynurenine to 3-hydroxykynurenine. KMO has a key role in tryptophan catabolism and synthesis of ommochrome pigments in mosquitoes. The gene encoding this enzyme in the yellow fever mosquito, Aedes aegypti, is called kynurenine hydroxylase (kh) and a mutant allele that produces white eyes has been designated khw. A number of cDNA clones representative of wild-type and mutant genes were isolated. Sequence analyses of the wild-type and mutant cDNAs revealed a deletion of 162 nucleotides in the mutant gene near the 3'-end of the deduced coding region. RT-PCR analyses confirm the transcription of a truncated mRNA in the mutant strain. The in-frame deletion results in a loss of 54 amino acids, which disrupts a major α-helix and which probably accounts for the loss of activity of the enzyme. Recombinant Ae. aegypti KMO showed high substrate specificity for kynurenine with optimum activity at 40 °C and pH = 7.5. Kinetic parameters and inhibition of KMO activity by Cl- and pyridoxal-5-phosphate were determined.</description><subject>3-hydroxykynurenine</subject><subject>Aedes - enzymology</subject><subject>Aedes - genetics</subject><subject>Aedes aegypti</subject><subject>amino acid sequences</subject><subject>Animals</subject><subject>complementary DNA</subject><subject>Culicidae</subject><subject>deletion mutants</subject><subject>DNA, Complementary - genetics</subject><subject>DNA, Complementary - isolation & purification</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>enzyme activity</subject><subject>enzyme inhibition</subject><subject>enzyme kinetics</subject><subject>Gene Deletion</subject><subject>gene expression</subject><subject>Gene Expression Profiling</subject><subject>insect vectors</subject><subject>Kinetics</subject><subject>KMO gene</subject><subject>Kynurenine</subject><subject>Kynurenine 3-Monooxygenase</subject><subject>kynurenine hydroxylase</subject><subject>messenger RNA</subject><subject>Mixed Function Oxygenases - chemistry</subject><subject>Mixed Function Oxygenases - genetics</subject><subject>molecular sequence data</subject><subject>mutants</subject><subject>mutation</subject><subject>nucleotide sequences</subject><subject>open reading frames</subject><subject>oxygenases</subject><subject>recombinant proteins</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Sequence Alignment</subject><subject>structural genes</subject><subject>substrate specificity</subject><subject>truncation mutants</subject><subject>white eye</subject><subject>Yellow fever virus</subject><issn>0962-1075</issn><issn>1365-2583</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkV1v0zAUhiMEYmXwF8BXXC3FH4kTSwipm9iYKEN8TFweuclJ5y6xuzjZGm7547hLKXAFVz7Sed5XPnqiiDA6ZTSRr1ZTJmQa8zQXU06pmFKaCDHdPIgm-8XDaEKV5DGjWXoQPfF-RSnNlVSPowPGVZaoVEyiHzOr68EbT1xFuiskd6Yu425YI9G2JE3faduRJVr0BG3hSmOX9xza70OD5HqwfYvWWCSNs85thsBqj7_qBqxrd0cqvMU2EP6mN507IjMsQ6HG5bDuzNPoUaVrj89272F0efr268m7eP7x7PxkNo8LyXMRl1mRpbmsKFU8wQXlPGGYF6ISmjHOUNKiLJXMlc4qhkUicCEKXWIp5EInVIvD6M3Yu-4XDZYF2q7VNaxb0-h2AKcN_L2x5gqW7ha45CpVLBS83BW07qZH30FjfBEu1BZd7yETMpCM_xNkuWJMijSA-QgWrfO-xWr_G0ZhqxpWsDUKW6OwVQ33qmETos__vOZ3cOc2AK9HICjF4b-L4fzDcRhCPB7jxne42cd1ew0yE1kK3y7OgL__PGenx5_gIvAvRr7SDvSyNR4uv3DKRPCV0owq8RPvptPg</recordid><startdate>200310</startdate><enddate>200310</enddate><creator>Han, Q</creator><creator>Calvo, E</creator><creator>Marinotti, O</creator><creator>Fang, J</creator><creator>Rizzi, M</creator><creator>James, A.A</creator><creator>Li, J</creator><general>Blackwell Science Ltd</general><scope>FBQ</scope><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7SS</scope><scope>8FD</scope><scope>C1K</scope><scope>F1W</scope><scope>FR3</scope><scope>H95</scope><scope>H97</scope><scope>L.G</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>200310</creationdate><title>Analysis of the wild-type and mutant genes encoding the enzyme kynurenine monooxygenase of the yellow fever mosquito, Aedes aegypti</title><author>Han, Q ; Calvo, E ; Marinotti, O ; Fang, J ; Rizzi, M ; James, A.A ; Li, J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c6283-d7c7586f00924eb02241e8c3f3a1121e60cdd9689a7f1ec43eb3caded36ba40a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>3-hydroxykynurenine</topic><topic>Aedes - enzymology</topic><topic>Aedes - genetics</topic><topic>Aedes aegypti</topic><topic>amino acid sequences</topic><topic>Animals</topic><topic>complementary DNA</topic><topic>Culicidae</topic><topic>deletion mutants</topic><topic>DNA, Complementary - genetics</topic><topic>DNA, Complementary - isolation & purification</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>enzyme activity</topic><topic>enzyme inhibition</topic><topic>enzyme kinetics</topic><topic>Gene Deletion</topic><topic>gene expression</topic><topic>Gene Expression Profiling</topic><topic>insect vectors</topic><topic>Kinetics</topic><topic>KMO gene</topic><topic>Kynurenine</topic><topic>Kynurenine 3-Monooxygenase</topic><topic>kynurenine hydroxylase</topic><topic>messenger RNA</topic><topic>Mixed Function Oxygenases - chemistry</topic><topic>Mixed Function Oxygenases - genetics</topic><topic>molecular sequence data</topic><topic>mutants</topic><topic>mutation</topic><topic>nucleotide sequences</topic><topic>open reading frames</topic><topic>oxygenases</topic><topic>recombinant proteins</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Sequence Alignment</topic><topic>structural genes</topic><topic>substrate specificity</topic><topic>truncation mutants</topic><topic>white eye</topic><topic>Yellow fever virus</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Han, Q</creatorcontrib><creatorcontrib>Calvo, E</creatorcontrib><creatorcontrib>Marinotti, O</creatorcontrib><creatorcontrib>Fang, J</creatorcontrib><creatorcontrib>Rizzi, M</creatorcontrib><creatorcontrib>James, A.A</creatorcontrib><creatorcontrib>Li, J</creatorcontrib><collection>AGRIS</collection><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Engineering Research Database</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 3: Aquatic Pollution & Environmental Quality</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Insect molecular biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Han, Q</au><au>Calvo, E</au><au>Marinotti, O</au><au>Fang, J</au><au>Rizzi, M</au><au>James, A.A</au><au>Li, J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Analysis of the wild-type and mutant genes encoding the enzyme kynurenine monooxygenase of the yellow fever mosquito, Aedes aegypti</atitle><jtitle>Insect molecular biology</jtitle><addtitle>Insect Mol Biol</addtitle><date>2003-10</date><risdate>2003</risdate><volume>12</volume><issue>5</issue><spage>483</spage><epage>490</epage><pages>483-490</pages><issn>0962-1075</issn><eissn>1365-2583</eissn><abstract>Kynurenine 3-monooxygenase (KMO) catalyses the hydroxylation of kynurenine to 3-hydroxykynurenine. KMO has a key role in tryptophan catabolism and synthesis of ommochrome pigments in mosquitoes. The gene encoding this enzyme in the yellow fever mosquito, Aedes aegypti, is called kynurenine hydroxylase (kh) and a mutant allele that produces white eyes has been designated khw. A number of cDNA clones representative of wild-type and mutant genes were isolated. Sequence analyses of the wild-type and mutant cDNAs revealed a deletion of 162 nucleotides in the mutant gene near the 3'-end of the deduced coding region. RT-PCR analyses confirm the transcription of a truncated mRNA in the mutant strain. The in-frame deletion results in a loss of 54 amino acids, which disrupts a major α-helix and which probably accounts for the loss of activity of the enzyme. Recombinant Ae. aegypti KMO showed high substrate specificity for kynurenine with optimum activity at 40 °C and pH = 7.5. Kinetic parameters and inhibition of KMO activity by Cl- and pyridoxal-5-phosphate were determined.</abstract><cop>Oxford, UK</cop><pub>Blackwell Science Ltd</pub><pmid>12974953</pmid><doi>10.1046/j.1365-2583.2003.00433.x</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Insect molecular biology, 2003-10, Vol.12 (5), p.483-490 |
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| subjects | 3-hydroxykynurenine Aedes - enzymology Aedes - genetics Aedes aegypti amino acid sequences Animals complementary DNA Culicidae deletion mutants DNA, Complementary - genetics DNA, Complementary - isolation & purification Electrophoresis, Polyacrylamide Gel enzyme activity enzyme inhibition enzyme kinetics Gene Deletion gene expression Gene Expression Profiling insect vectors Kinetics KMO gene Kynurenine Kynurenine 3-Monooxygenase kynurenine hydroxylase messenger RNA Mixed Function Oxygenases - chemistry Mixed Function Oxygenases - genetics molecular sequence data mutants mutation nucleotide sequences open reading frames oxygenases recombinant proteins Reverse Transcriptase Polymerase Chain Reaction Sequence Alignment structural genes substrate specificity truncation mutants white eye Yellow fever virus |
| title | Analysis of the wild-type and mutant genes encoding the enzyme kynurenine monooxygenase of the yellow fever mosquito, Aedes aegypti |
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