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Extensin Peroxidase Identification a...

Dong, Wen.

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Extensin Peroxidase Identification and Characterization in Solanum lycopersicum.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Extensin Peroxidase Identification and Characterization in Solanum lycopersicum./
作者:	Dong, Wen.
面頁冊數:	205 p.
附註:	Source: Dissertation Abstracts International, Volume: 77-02(E), Section: B.
Contained By:	Dissertation Abstracts International77-02B(E).
標題:	Biochemistry. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3729408
ISBN:	9781339152622

Extensin Peroxidase Identification and Characterization in Solanum lycopersicum.
Dong, Wen.

Extensin Peroxidase Identification and Characterization in Solanum lycopersicum. - 205 p.

Source: Dissertation Abstracts International, Volume: 77-02(E), Section: B.

Thesis (Ph.D.)--Ohio University, 2015.

The regulation of plant cell growth and defense involves the insolubilization of hydroxyproline-rich glycoproteins (HRGPs), such as extensin, in the primary cell wall. In tomato (Solanum lycopersicum ), insolublization occurs by the formation of tyrosylcrosslinks catalyzed specifically by the pI 4.6 extensin peroxidase (EP). To date, neither the gene encoding EP nor the protein itself has been identified.

ISBN: 9781339152622Subjects--Topical Terms:

518028
Biochemistry.

Extensin Peroxidase Identification and Characterization in Solanum lycopersicum.
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100 1 $a Dong, Wen. $3 3181910
245 1 0 $a Extensin Peroxidase Identification and Characterization in Solanum lycopersicum.
300 $a 205 p.
500 $a Source: Dissertation Abstracts International, Volume: 77-02(E), Section: B.
500 $a Advisers: Marcia J. Kieliszewski; Michael A. Held.
502 $a Thesis (Ph.D.)--Ohio University, 2015.
520 $a The regulation of plant cell growth and defense involves the insolubilization of hydroxyproline-rich glycoproteins (HRGPs), such as extensin, in the primary cell wall. In tomato (Solanum lycopersicum ), insolublization occurs by the formation of tyrosylcrosslinks catalyzed specifically by the pI 4.6 extensin peroxidase (EP). To date, neither the gene encoding EP nor the protein itself has been identified.
520 $a Here, tomato EP candidates were identified using both proteomic and bioinformatic approaches. Bioinformatic screening of the tomato genome yielded eight EP gene candidates, which encoded a putative signal sequence and a protein with a predicted pI near 4.6. Biochemical fractionation of tomato culture media followed by proteomic detection further refined our list of EP candidates to three, with the lead candidate designated a "CG5". The CG5 open-reading frame from a tomato cDNA was then cloned into a bacterial expression vector to test for EP crosslinking activity.
520 $a CG5 was expressed in E. coli, fractionated from inclusion bodies, and folded in vitro. Peroxidase activity of CG5 was assayed and quantified by the ABTS (2,2'- zinobis(3-ethylbenzothiazoline-6-sulphonic acid)) assay. Subsequent extensin crosslinking assays showed that CG5 can covalently crosslink authentic tomato P1 extensin and P3-type extensin analogs in vitro supporting the hypothesis that CG5 encodes a tomato EP. Recombinant CG5 was then purified for enzyme kinetics study which gave out a Km of 0.105 mM. A computational 3D model was generated for CG5 with highlighted active site and predicted heme group binding.
590 $a School code: 0167.
650 4 $a Biochemistry. $3 518028
650 4 $a Botany. $3 516217
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710 2 $a Ohio University. $b Chemistry and Biochemistry (Arts and Sciences). $3 3181911
773 0 $t Dissertation Abstracts International $g 77-02B(E).
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792 $a 2015
793 $a English
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