Material Information |
| Title: |
Kinetic Characterization of Caenorhabditis elegans postembryonic Glyceraldehyde-3-Phosphate Dehydrogenase |
| Physical Description: |
Book |
| Language: |
English |
| Creator: |
Silimon, Ruth |
| Publisher: |
New College of Florida |
| Place of Publication: |
Sarasota, Fla. |
| Creation Date: |
2013 |
| Publication Date: |
2013 |
Subjects |
| Subjects / Keywords: |
Kinetics
C. Elegans
Glyceraldehyde-3-Phosphate Dehydrogenase |
| Genre: |
bibliography ( marcgt )
theses ( marcgt )
government publication (state, provincial, terriorial, dependent) ( marcgt )
born-digital ( sobekcm )
Electronic Thesis or Dissertation |
Notes |
| Abstract: |
Caenorhabditis elegans is a model organism in modern biological research. The worm has four different genes, gpd 1-4, for the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). GAPDH catalyzes the oxidation and phosphorylation of glyceraldehyde-3-phosphate (G3P) to 1,3-bisphosphoglycerate (1,3-BPG) using NAD+ and Pi as substrates. The 1,3-BPG product is used in a later reaction in glycolysis to generate ATP. The physiological significance of multiple GAPDH isoenzymes within C. elegans is still not understood. This is the first project to characterize partially purified GPD-3 formally. The IMPACT purification system from NEB was used in conjunction with Rosetta (DE3) cell hosts to over-express and partially purify the enzyme. Enzyme kinetics were performed with all three substrates at pH 8.0. The Km values for NAD+, G3P, and Pi were estimated to be 0.3 mM, 2.6 mM, and 0.4 mM, respectively. The enzyme's specific activity was found to be within the range of 3.6 to 5.3 µmol/min•mg. The Km for NAD+ reported here is lower than the value found for an endogenous GPD mixture however, the Km for G3P is much higher, suggesting the need for further analysis. |
| Statement of Responsibility: |
by Ruth Silimon |
| Thesis: |
Thesis (B.A.) -- New College of Florida, 2013 |
| General Note: |
Online version not currently available. |
| Electronic Access: |
RESTRICTED TO NCF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE |
| Bibliography: |
Includes bibliographical references. |
| Source of Description: |
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. |
| Local: |
Faculty Sponsor: Walstrom, Katherine |
Record Information |
| Source Institution: |
New College of Florida |
| Holding Location: |
New College of Florida |
| Rights Management: |
Applicable rights reserved. |
| Classification: |
local - S.T. 2013 S58 |
| System ID: |
NCFE004865:00001 |
|
Material Information |
| Title: |
Kinetic Characterization of Caenorhabditis elegans postembryonic Glyceraldehyde-3-Phosphate Dehydrogenase |
| Physical Description: |
Book |
| Language: |
English |
| Creator: |
Silimon, Ruth |
| Publisher: |
New College of Florida |
| Place of Publication: |
Sarasota, Fla. |
| Creation Date: |
2013 |
| Publication Date: |
2013 |
Subjects |
| Subjects / Keywords: |
Kinetics
C. Elegans
Glyceraldehyde-3-Phosphate Dehydrogenase |
| Genre: |
bibliography ( marcgt )
theses ( marcgt )
government publication (state, provincial, terriorial, dependent) ( marcgt )
born-digital ( sobekcm )
Electronic Thesis or Dissertation |
Notes |
| Abstract: |
Caenorhabditis elegans is a model organism in modern biological research. The worm has four different genes, gpd 1-4, for the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). GAPDH catalyzes the oxidation and phosphorylation of glyceraldehyde-3-phosphate (G3P) to 1,3-bisphosphoglycerate (1,3-BPG) using NAD+ and Pi as substrates. The 1,3-BPG product is used in a later reaction in glycolysis to generate ATP. The physiological significance of multiple GAPDH isoenzymes within C. elegans is still not understood. This is the first project to characterize partially purified GPD-3 formally. The IMPACT purification system from NEB was used in conjunction with Rosetta (DE3) cell hosts to over-express and partially purify the enzyme. Enzyme kinetics were performed with all three substrates at pH 8.0. The Km values for NAD+, G3P, and Pi were estimated to be 0.3 mM, 2.6 mM, and 0.4 mM, respectively. The enzyme's specific activity was found to be within the range of 3.6 to 5.3 µmol/min•mg. The Km for NAD+ reported here is lower than the value found for an endogenous GPD mixture however, the Km for G3P is much higher, suggesting the need for further analysis. |
| Statement of Responsibility: |
by Ruth Silimon |
| Thesis: |
Thesis (B.A.) -- New College of Florida, 2013 |
| General Note: |
Online version not currently available. |
| Electronic Access: |
RESTRICTED TO NCF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE |
| Bibliography: |
Includes bibliographical references. |
| Source of Description: |
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. |
| Local: |
Faculty Sponsor: Walstrom, Katherine |
Record Information |
| Source Institution: |
New College of Florida |
| Holding Location: |
New College of Florida |
| Rights Management: |
Applicable rights reserved. |
| Classification: |
local - S.T. 2013 S58 |
| System ID: |
NCFE004865:00001 |
|