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PROBING PROTEOMES WITH ALTERNATIVE PROTEASES: LYS-N & ARG-C

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Material Information

Title:
PROBING PROTEOMES WITH ALTERNATIVE PROTEASES: LYS-N & ARG-C
Physical Description:
Book
Language:
English
Creator:
Platé IV, Thomas Earle
Publisher:
New College of Florida
Place of Publication:
Sarasota, Fla.
Publication Date:

Thesis/Dissertation Information

Degree:
Bachelor's ( B.A.)
Degree Grantor:
New College of Florida
Degree Divisions:
Natural Sciences
Area of Concentration:
Biochemistry
Faculty Sponsor:
Scudder, Paul

Subjects

Genre:
Electronic Thesis or Dissertation
bibliography   ( marcgt )
theses   ( marcgt )

Notes

Abstract:
Proteomics, the study of the protein functional complement of the genome, deals with similar issues of amino-acid sequence coverage and depth. Trypsin has been the most common protease used to cleave peptides into manageable units due the many optimum traits of tryptic digestions for mass spectrometry (MS) analyses for the entire history of “shotgun” proteomics. Instrumental and computational methods for proteomic analyses have been tailored for tryptic peptides as a result. This project explored the feasibility of an alternative approach that utilizes a sequential digestion by lesser-used proteases, a C-terminal arginine peptidase (Arg-C or Clostripain) and a specific N-terminal lysine metallopeptidase (Lys-N). Lys-N was isolated from Grifola frondosa (maitake), and Arg-C was obtained from a commercial source. Doubly-digested peptides from a mouse brain protein extract were separated by offline strong cation exchange (SCX) chromatography followed by on-line ultra-performance liquid chromatography (UPLC) and analysis by high resolution mass spectrometry. This preparation produced a population of unique peptides that were tightly retained on the SCX column and ionized well by electrospray ionization (ESI). Peptides were identifiable via extracted ion tracking as well as the SEQUEST search algorithm. Thus, the method described herein represents a potential alternate route to acquire complementary data to conventional proteomic approaches.
Thesis:
Thesis (B.A.) -- New College of Florida, 2014
General Note:
Accompanying materials: CD containing database search results.
General Note:
RESTRICTED TO NCF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE
Bibliography:
Includes bibliographical references.
General Note:
This bibliographic record is available under the Creative Commons CC0 public domain dedication. The New College of Florida Libraries, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
General Note:
Faculty Sponsor: Scudder, Paul
Statement of Responsibility:
by Thomas Earle Platé IV

Record Information

Source Institution:
New College of Florida
Holding Location:
New College of Florida
Rights Management:
Applicable rights reserved.
Resource Identifier:
Classification:
S.T. 2014 P5
System ID:
AA00024701:00001

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