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Acyl-CoA synthetases in Trypanosoma ...

Jiang, David Wei.

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Acyl-CoA synthetases in Trypanosoma brucei.

紀錄類型:	書目-語言資料,印刷品 : Monograph/item
正題名/作者:	Acyl-CoA synthetases in Trypanosoma brucei./
作者:	Jiang, David Wei.
面頁冊數:	129 p.
附註:	Adviser: Paul Englund.
Contained By:	Dissertation Abstracts International62-10B.
標題:	Biology, Microbiology. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3028286
ISBN:	049340337X

Acyl-CoA synthetases in Trypanosoma brucei.
Jiang, David Wei.

Acyl-CoA synthetases in Trypanosoma brucei. - 129 p.

Adviser: Paul Englund.

Thesis (Ph.D.)--The Johns Hopkins University, 2002.

Trypanosomes are single-celled flagellated protozoa that infect a wide variety of hosts. They evade the host immune response by undergoing antigenic variation, which is mediated by a surface coat composed of 10<super>7</super> identical variant surface glycoprotein (VSG) molecules. The VSG's C-terminal amino acid is linked to a glycosyl phosphatidylinositol membrane anchor. An important feature of the bloodstream form trypanosome GPI anchor is the exclusive presence of two myristates, a 14 carbon saturated fatty acid. One key step in the processing of myristate for incorporation into GPIs is the conversion of myristate to myristoyl-CoA, a reaction catalyzed by an acyl-CoA synthetase.

ISBN: 049340337XSubjects--Topical Terms:

1017734
Biology, Microbiology.

Acyl-CoA synthetases in Trypanosoma brucei.
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100 1 $a Jiang, David Wei. $3 1251207
245 1 0 $a Acyl-CoA synthetases in Trypanosoma brucei.
300 $a 129 p.
500 $a Adviser: Paul Englund.
500 $a Source: Dissertation Abstracts International, Volume: 62-10, Section: B, page: 4526.
502 $a Thesis (Ph.D.)--The Johns Hopkins University, 2002.
520 $a Trypanosomes are single-celled flagellated protozoa that infect a wide variety of hosts. They evade the host immune response by undergoing antigenic variation, which is mediated by a surface coat composed of 10<super>7</super> identical variant surface glycoprotein (VSG) molecules. The VSG's C-terminal amino acid is linked to a glycosyl phosphatidylinositol membrane anchor. An important feature of the bloodstream form trypanosome GPI anchor is the exclusive presence of two myristates, a 14 carbon saturated fatty acid. One key step in the processing of myristate for incorporation into GPIs is the conversion of myristate to myristoyl-CoA, a reaction catalyzed by an acyl-CoA synthetase.
520 $a I investigated three aspects of <italic>Trypanosoma brucei</italic> acyl-CoA synthetase (TbACS). First, I have cloned four tandemly linked TbACS genes, which reside in a 12.6 kb region of the genome. These genes are present in a single copy, and all four are expressed in both bloodstream and procyclic trypanosomes. Sequence alignments indicate that the encoded proteins are 46%–95% identical. Three of them share almost identical C-termini.
520 $a Second, I have expressed these four genes in <italic>Escherichia coli </italic> and purified the products to near homogeneity. Although these enzymes are highly homologous, they have distinct specificities for fatty acid chain length. TbACS1 prefers saturated fatty acids in the range of C11:0 to C14:0, and TbACS2 prefers shorter fatty acids, mainly C10:0. TbACS3 and 4, which have 95% sequence identity, have similar specificities, favoring fatty acids between C14:0 and C17:0. In addition, TbACS1, 3, and 4 function well with a variety of unsaturated fatty acids.
520 $a Furthermore, I have purified and identified a major TbACS from bloodstream form trypanosomes. The enzyme was extracted with 0.1% Triton X-100 and purified using Q-Sepharose column chromatography. Only one band at 78 kDa on the SDS-PAGE gel correlates with the ACS activity. In-gel trypsin digestion and MALDI-TOF mass spectrometry analysis revealed that the band contained a mixture of TbACS3 and BiP, an abundant chaperone protein. The partially purified TbACS3 had substrate specificity similar to that of the recombinant TbACS3.
590 $a School code: 0098.
650 4 $a Biology, Microbiology. $3 1017734
650 4 $a Biology, Molecular. $3 1017719
650 4 $a Chemistry, Biochemistry. $3 1017722
690 $a 0307
690 $a 0410
690 $a 0487
710 2 0 $a The Johns Hopkins University. $3 1017431
773 0 $t Dissertation Abstracts International $g 62-10B.
790 $a 0098
790 1 0 $a Englund, Paul, $e advisor
791 $a Ph.D.
792 $a 2002
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3028286