東華大學圖書館 |

Language: English

Help

回圖書館首頁

手機版館藏查詢

Back

Switch To: Labeled | MARC Mode | ISBD

CryoEM-Enabled Approaches to Structu...

Ho, Chi-Min.

Linked to FindBook

Google Book

Amazon

博客來

CryoEM-Enabled Approaches to Structure Determination of Endogenous Protein Complexes Implicated in the Pathogenesis of the Malaria Parasite Plasmodium falciparum.

Record Type:	Electronic resources : Monograph/item
Title/Author:	CryoEM-Enabled Approaches to Structure Determination of Endogenous Protein Complexes Implicated in the Pathogenesis of the Malaria Parasite Plasmodium falciparum./
Author:	Ho, Chi-Min.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2019,
Description:	151 p.
Notes:	Source: Dissertations Abstracts International, Volume: 80-10, Section: B.
Contained By:	Dissertations Abstracts International80-10B.
Subject:	Biochemistry. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=13806796
ISBN:	9781392011300

CryoEM-Enabled Approaches to Structure Determination of Endogenous Protein Complexes Implicated in the Pathogenesis of the Malaria Parasite Plasmodium falciparum.
Ho, Chi-Min.

CryoEM-Enabled Approaches to Structure Determination of Endogenous Protein Complexes Implicated in the Pathogenesis of the Malaria Parasite Plasmodium falciparum. - Ann Arbor : ProQuest Dissertations & Theses, 2019 - 151 p.

Source: Dissertations Abstracts International, Volume: 80-10, Section: B.

Thesis (Ph.D.)--University of California, Los Angeles, 2019.

This item is not available from ProQuest Dissertations & Theses.

The complexity and breadth of the host-cell remodeling machinery in the malaria parasite P. falciparum make it a rich and exciting system for the study of host-pathogen interfaces, particularly as many of the molecular mechanisms underlying this parasite's ability to hijack human red blood cells remain unclear. Furthermore, the P. falciparum proteome has proven recalcitrant to structural and biochemical characterization using recombinant methods, making it an intriguing model system for the development of new methods that leverage recent advances in cryoEM to enable structural studies of previously intractable systems at near-atomic resolution. The work presented here makes significant contributions in both these regards. First, we use a targeted, CRISPR-enabled "top down" approach to determine near-atomic resolution structures of the unique malaria parasite translocon PTEX, which we purified directly from P. falciparum parasites in multiple functional states, yielding the first near-atomic resolution cryoEM structures of a protein isolated directly from an endogenous source using an epitope tag inserted into the endogenous locus with CRISPR-Cas9 gene editing. We then developed a "bottom up" endogenous structural proteomics method whereby protein complexes enriched directly from the cellular milieu are identified by imaging and structure determination using cryoEM and mass spectrometry. As a proof of principle, we successfully used this approach to obtain near-atomic resolution structures of multiple protein complexes from the P. falciparum proteome, which has previously proven recalcitrant to expression in recombinant systems, precluding structure determination by X-ray crystallography or NMR. The body of work described here addresses a known need for methods that overcome the limitations of structural biology approaches that depend on recombinant systems, opening the door for high resolution structure determination of a vast number of previously intractable biological systems.

ISBN: 9781392011300Subjects--Topical Terms:

518028
Biochemistry.
Subjects--Index Terms:

CryoEM

CryoEM-Enabled Approaches to Structure Determination of Endogenous Protein Complexes Implicated in the Pathogenesis of the Malaria Parasite Plasmodium falciparum.
LDR:03493nmm a2200421 4500 001 2272385
005 20201105110042.5
008 220629s2019 ||||||||||||||||| ||eng d
020 $a 9781392011300
035 $a (MiAaPQ)AAI13806796
035 $a (MiAaPQ)ucla:17542
035 $a AAI13806796
040 $a MiAaPQ $c MiAaPQ
100 1 $a Ho, Chi-Min. $3 3549820
245 1 0 $a CryoEM-Enabled Approaches to Structure Determination of Endogenous Protein Complexes Implicated in the Pathogenesis of the Malaria Parasite Plasmodium falciparum.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2019
300 $a 151 p.
500 $a Source: Dissertations Abstracts International, Volume: 80-10, Section: B.
500 $a Publisher info.: Dissertation/Thesis.
500 $a Includes supplementary digital materials.
500 $a Advisor: Zhou, Hong.
502 $a Thesis (Ph.D.)--University of California, Los Angeles, 2019.
506 $a This item is not available from ProQuest Dissertations & Theses.
506 $a This item must not be sold to any third party vendors.
520 $a The complexity and breadth of the host-cell remodeling machinery in the malaria parasite P. falciparum make it a rich and exciting system for the study of host-pathogen interfaces, particularly as many of the molecular mechanisms underlying this parasite's ability to hijack human red blood cells remain unclear. Furthermore, the P. falciparum proteome has proven recalcitrant to structural and biochemical characterization using recombinant methods, making it an intriguing model system for the development of new methods that leverage recent advances in cryoEM to enable structural studies of previously intractable systems at near-atomic resolution. The work presented here makes significant contributions in both these regards. First, we use a targeted, CRISPR-enabled "top down" approach to determine near-atomic resolution structures of the unique malaria parasite translocon PTEX, which we purified directly from P. falciparum parasites in multiple functional states, yielding the first near-atomic resolution cryoEM structures of a protein isolated directly from an endogenous source using an epitope tag inserted into the endogenous locus with CRISPR-Cas9 gene editing. We then developed a "bottom up" endogenous structural proteomics method whereby protein complexes enriched directly from the cellular milieu are identified by imaging and structure determination using cryoEM and mass spectrometry. As a proof of principle, we successfully used this approach to obtain near-atomic resolution structures of multiple protein complexes from the P. falciparum proteome, which has previously proven recalcitrant to expression in recombinant systems, precluding structure determination by X-ray crystallography or NMR. The body of work described here addresses a known need for methods that overcome the limitations of structural biology approaches that depend on recombinant systems, opening the door for high resolution structure determination of a vast number of previously intractable biological systems.
590 $a School code: 0031.
650 4 $a Biochemistry. $3 518028
650 4 $a Biophysics. $3 518360
653 $a CryoEM
653 $a CryoID
653 $a Malaria
653 $a Membrane protein
653 $a Structural proteomics
653 $a Translocon
690 $a 0487
690 $a 0786
710 2 $a University of California, Los Angeles. $b Molecular Biology. $3 3283994
773 0 $t Dissertations Abstracts International $g 80-10B.
790 $a 0031
791 $a Ph.D.
792 $a 2019
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=13806796