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Liu, Fan.

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The Development and Application of Novel Crosslinking Mass Spectrometry Approaches to Study Protein Structure and Interactions.

Record Type:	Language materials, printed : Monograph/item
Title/Author:	The Development and Application of Novel Crosslinking Mass Spectrometry Approaches to Study Protein Structure and Interactions./
Author:	Liu, Fan.
Description:	203 p.
Notes:	Source: Dissertation Abstracts International, Volume: 75-03(E), Section: B.
Contained By:	Dissertation Abstracts International75-03B(E).
Subject:	Chemistry, Biochemistry. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3575785
ISBN:	9781303547669

The Development and Application of Novel Crosslinking Mass Spectrometry Approaches to Study Protein Structure and Interactions.
Liu, Fan.

The Development and Application of Novel Crosslinking Mass Spectrometry Approaches to Study Protein Structure and Interactions. - 203 p.

Source: Dissertation Abstracts International, Volume: 75-03(E), Section: B.

Thesis (Ph.D.)--North Carolina State University, 2013.

Chemical crosslinking combined with mass spectrometry (XL-MS) is a powerful approach to investigate protein conformation and protein-protein interactions. The length of a given crosslinker provides a distance constraint which is useful to reveal structural information on proteins (intra-protein crosslinks) and interacting partners (inter-protein crosslinks). However, the identification of the residues involved in the crosslink remains analytically challenging due to 1) the complexity of the sample (myriad of crosslinked products present at various stoichiometries where the most informative inter-peptide crosslinks are among the lowest abundance) and 2) the complexity of the MS 2 spectrum (the precursor mass obtained for inter-peptide crosslink is the sum of the two linked peptides as opposed to each individual peptide). To address these challenges, we have developed combinatorial gas-phase cleavable crosslinkers (disuccinimidyl-succinamyl-aspartyl-proline (SuDP) and disuccinimidyl-succinamyl-valyl-proline (SuVP)) (Chapter 2), a series of LC/MSn strategies (the two-step LC/MSn and the LC/MS2 only approaches) and crosslink search engines (CXLinkS and CXLink) (Chapters 3 and 4, respectively) to enable robust and reliable inter-peptide crosslink identification from large proteome databases. We verified our approaches with standard proteins (bovine serum albumin and human hemoglobin) and a large protein complex (E. coli 30S ribosome) and then applied them to unravel the assembly of Bacillus subtilis transition state regulator AbrB tetramer (Chapter 5) and the interaction of Arabidopsis thaliana BRASSINOSTEROID INSENSITIVE 1 (BRI1) and BRI1 ASSOCIATED RECEPTOR KINASE 1 (BAK1) (chapter 6). Overall, we successfully developed a series of XL-MS strategies (including novel crosslinkers and software) and lays the foundation for high-throughput investigations of protein complex structures and interactome configurations.

ISBN: 9781303547669Subjects--Topical Terms:

1017722
Chemistry, Biochemistry.

The Development and Application of Novel Crosslinking Mass Spectrometry Approaches to Study Protein Structure and Interactions.
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245 1 4 $a The Development and Application of Novel Crosslinking Mass Spectrometry Approaches to Study Protein Structure and Interactions.
300 $a 203 p.
500 $a Source: Dissertation Abstracts International, Volume: 75-03(E), Section: B.
500 $a Adviser: Michael Goshe.
502 $a Thesis (Ph.D.)--North Carolina State University, 2013.
520 $a Chemical crosslinking combined with mass spectrometry (XL-MS) is a powerful approach to investigate protein conformation and protein-protein interactions. The length of a given crosslinker provides a distance constraint which is useful to reveal structural information on proteins (intra-protein crosslinks) and interacting partners (inter-protein crosslinks). However, the identification of the residues involved in the crosslink remains analytically challenging due to 1) the complexity of the sample (myriad of crosslinked products present at various stoichiometries where the most informative inter-peptide crosslinks are among the lowest abundance) and 2) the complexity of the MS 2 spectrum (the precursor mass obtained for inter-peptide crosslink is the sum of the two linked peptides as opposed to each individual peptide). To address these challenges, we have developed combinatorial gas-phase cleavable crosslinkers (disuccinimidyl-succinamyl-aspartyl-proline (SuDP) and disuccinimidyl-succinamyl-valyl-proline (SuVP)) (Chapter 2), a series of LC/MSn strategies (the two-step LC/MSn and the LC/MS2 only approaches) and crosslink search engines (CXLinkS and CXLink) (Chapters 3 and 4, respectively) to enable robust and reliable inter-peptide crosslink identification from large proteome databases. We verified our approaches with standard proteins (bovine serum albumin and human hemoglobin) and a large protein complex (E. coli 30S ribosome) and then applied them to unravel the assembly of Bacillus subtilis transition state regulator AbrB tetramer (Chapter 5) and the interaction of Arabidopsis thaliana BRASSINOSTEROID INSENSITIVE 1 (BRI1) and BRI1 ASSOCIATED RECEPTOR KINASE 1 (BAK1) (chapter 6). Overall, we successfully developed a series of XL-MS strategies (including novel crosslinkers and software) and lays the foundation for high-throughput investigations of protein complex structures and interactome configurations.
590 $a School code: 0155.
650 4 $a Chemistry, Biochemistry. $3 1017722
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