To fix, comment below in the following format:MSc http://purl.org/ontology/bibo/degrees/ms
Record:
=LDR 04373cam a2200373 a 4500
=001 3258426
=005 20160427150326.0
=006 m\\f\\d\\\\
=007 he\amu\\buua
=008 090707s2009\\kina\\smb\\000\0\eng\d
=040 \$aCaOKQ$beng$cCaOKQ
=100 1\$aWu, John.
=245 10$aIdentification of genes involved in the assembly and biosynthesis of the N-linked flagellin glycan in the archaeon, methanococcus maripaludis /$cby John Wu.
=264 \1$aKingston, Ont. :$b[publisher not identified],$c2009.
=300 \$aix, 15 leaves :$billustrations (some color)
=336 \$atext$btxt$2rdacontent
=337 \$acomputer$bc$2rdamedia
=338 \$aonline resource$bcr$2rdacarrier
=502 \$aMaster, Microbiology & Immunology$bQueen's University$c2009
=504 \$aIncludes bibliographical references (leaves 96-106)
=520 3\$aN-glycosylation is a metabolic process found in all three domains of life. It is the attachment of a polysaccharide glycan to asparagine (Asn) residues within the amino acid motif, Asn-Xaa-Ser/Thr. In the archaeon, Methanococcus maripaludis, a tetrasaccharide glycan was isolated from purified flagella and its structure determined by mass spectrometry analysis. The linking sugar to the protein is surprisingly, N-acetylgalactosamine (β-GalNAc), with the next proximal sugar a derivative of N-acetylglucosamine (β-GlcNAc), being named β-GlcNAc3Ac, and the third sugar a derivative of N-acetylmannosamine (β-ManNAc), with an attached threonine residue on the C6 carbon (β-ManNAc3NAm). The terminal sugar is an unusual diglycoside of aldulose ((5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose). Previous genetic analyses identified the glycosyltransferases (GTs) responsible for the transfer of the second and third sugars of the glycan, as well as the oligosaccharyltransferase (OST) which attaches the glycan to protein. Left unidentified were the first and fourth GTs, the flippase as well as any genes involved in glycan sugar biosynthesis and modification. In this work, genes suspected to be involved in the biosynthesis of N-linked sugars, as well as those that might encode the missing GTs and flippase were targeted for in-frame deletion. Mutants with a deleted annotated GT gene (MMP1088) had a small decrease in flagellin molecular weight as determined by immunoblotting. Mass spectrometry (MS) analysis confirmed that the N-linked glycan was missing the terminal sugar as well as the threonine found on the third sugar of wildtype cells. Mutants with a deleted gene annotated to be involved in acetamidino synthesis (a functional group that is present on the third sugar), also had a decrease in flagellin molecular weight. MS analysis determined that the N-linked glycan was missing the acetamidino group on the third sugar as well as its attached threonine, along with the terminal sugar. Both mutants were able to assemble functional flagella but had impaired motility compared to wildtype cells in mini-swarm agar. Deletions were also constructed in four other GT genes considered candidates in assembly of the linking sugar. However, none of these mutants had the expected decrease in flagellin molecular weight. With the work done in this study, the glycosyl transferase that attaches the last sugar of the M. maripaludis N-linked assembly pathway has been identified as well as a gene involved in the biosynthesis and modification of the glycan sugars.
=533 \$aCopy 2, microfiche.$bOttawa :$cNational Library of Canada,$d[2009] --$e2 microfiches ; 11 x 15 cm. --$f(Canadian theses = Thèses canadiennes)
=538 \$aMode of access: World Wide Web.
=540 \$aThis publication is made available by the authority of the copyright owner solely for the purpose of private study and research and may not be copied or reproduced except as permitted by the copyright laws without written authority from the copyright owner.
=500 \$aGMD: electronic resource.
=653 \$aN-glycosylation.
=653 \$aArchaea.
=653 \$aSugar biosynthesis.
=653 \$aGlycosyl transferase.
=710 2\$aQueen's University (Kingston, Ont.).$tTheses (Queen's University (Kingston, Ont.))
=710 2\$aQueen's University (Kingston, Ont.).$bDepartment of Microbiology and Immunology.
=830 \0$aCanadian theses.
=948 \$ac:sb
=902 \$aMARCIVE2013
The URI for Queen's University could not be found
To fix, comment below in the following format:
MSc http://purl.org/ontology/bibo/degrees/msRecord: =LDR 04373cam a2200373 a 4500 =001 3258426 =005 20160427150326.0 =006 m\\f\\d\\\\ =007 he\amu\\buua =008 090707s2009\\kina\\smb\\000\0\eng\d =040 \$aCaOKQ$beng$cCaOKQ =100 1\$aWu, John. =245 10$aIdentification of genes involved in the assembly and biosynthesis of the N-linked flagellin glycan in the archaeon, methanococcus maripaludis /$cby John Wu. =264 \1$aKingston, Ont. :$b[publisher not identified],$c2009. =300 \$aix, 15 leaves :$billustrations (some color) =336 \$atext$btxt$2rdacontent =337 \$acomputer$bc$2rdamedia =338 \$aonline resource$bcr$2rdacarrier =502 \$aMaster, Microbiology & Immunology$bQueen's University$c2009 =504 \$aIncludes bibliographical references (leaves 96-106) =520 3\$aN-glycosylation is a metabolic process found in all three domains of life. It is the attachment of a polysaccharide glycan to asparagine (Asn) residues within the amino acid motif, Asn-Xaa-Ser/Thr. In the archaeon, Methanococcus maripaludis, a tetrasaccharide glycan was isolated from purified flagella and its structure determined by mass spectrometry analysis. The linking sugar to the protein is surprisingly, N-acetylgalactosamine (β-GalNAc), with the next proximal sugar a derivative of N-acetylglucosamine (β-GlcNAc), being named β-GlcNAc3Ac, and the third sugar a derivative of N-acetylmannosamine (β-ManNAc), with an attached threonine residue on the C6 carbon (β-ManNAc3NAm). The terminal sugar is an unusual diglycoside of aldulose ((5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose). Previous genetic analyses identified the glycosyltransferases (GTs) responsible for the transfer of the second and third sugars of the glycan, as well as the oligosaccharyltransferase (OST) which attaches the glycan to protein. Left unidentified were the first and fourth GTs, the flippase as well as any genes involved in glycan sugar biosynthesis and modification. In this work, genes suspected to be involved in the biosynthesis of N-linked sugars, as well as those that might encode the missing GTs and flippase were targeted for in-frame deletion. Mutants with a deleted annotated GT gene (MMP1088) had a small decrease in flagellin molecular weight as determined by immunoblotting. Mass spectrometry (MS) analysis confirmed that the N-linked glycan was missing the terminal sugar as well as the threonine found on the third sugar of wildtype cells. Mutants with a deleted gene annotated to be involved in acetamidino synthesis (a functional group that is present on the third sugar), also had a decrease in flagellin molecular weight. MS analysis determined that the N-linked glycan was missing the acetamidino group on the third sugar as well as its attached threonine, along with the terminal sugar. Both mutants were able to assemble functional flagella but had impaired motility compared to wildtype cells in mini-swarm agar. Deletions were also constructed in four other GT genes considered candidates in assembly of the linking sugar. However, none of these mutants had the expected decrease in flagellin molecular weight. With the work done in this study, the glycosyl transferase that attaches the last sugar of the M. maripaludis N-linked assembly pathway has been identified as well as a gene involved in the biosynthesis and modification of the glycan sugars. =533 \$aCopy 2, microfiche.$bOttawa :$cNational Library of Canada,$d[2009] --$e2 microfiches ; 11 x 15 cm. --$f(Canadian theses = Thèses canadiennes) =538 \$aMode of access: World Wide Web. =540 \$aThis publication is made available by the authority of the copyright owner solely for the purpose of private study and research and may not be copied or reproduced except as permitted by the copyright laws without written authority from the copyright owner. =500 \$aGMD: electronic resource. =653 \$aN-glycosylation. =653 \$aArchaea. =653 \$aSugar biosynthesis. =653 \$aGlycosyl transferase. =710 2\$aQueen's University (Kingston, Ont.).$tTheses (Queen's University (Kingston, Ont.)) =710 2\$aQueen's University (Kingston, Ont.).$bDepartment of Microbiology and Immunology. =830 \0$aCanadian theses. =948 \$ac:sb =902 \$aMARCIVE2013
Record File: 1da4911d31d4a8313df26e3929f75498.mrc