Phadke M, Krynetskaia N, Mishra A, Barrero C, Merali S, Gothe SA, Krynetskiy E. Disruption of NAD+ binding site in glyceraldehyde 3-phosphate dehydrogenase affects its intranuclear interactions. World J Biol Chem 2015; 6(4): 366-378 [PMID: 26629320 DOI: 10.4331/wjbc.v6.i4.366]
Corresponding Author of This Article
Evgeny Krynetskiy, PhD, DSc, Associate Professor, Temple University School of Pharmacy, 3307 North Broad Street, Philadelphia, PA 19140, United States. ekrynets@temple.edu
Research Domain of This Article
Biochemistry & Molecular Biology
Article-Type of This Article
Basic Study
Open-Access Policy of This Article
This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Table 2 Kinetic parameters of wild type and mutated glyceraldehyde 3-phosphate dehydrogenase variants
GAPDH variant
Km (G3P) (μmol/L)
Km (NAD+) (μmol/L)
Wild type
77 ± 25.9
57 ± 11.1
Y94A
87 ± 33.1
97.4 ± 11
S98A
100 ± 36.7
101 ± 7.2
T99A
112 ± 44.7
105 ± 17.7
T99I
106 ± 38.3
741 ± 257
Table 3 Fluorescence recovery after photobleaching analysis of dynamic properties of wild type and mutated enhanced green fluorescent protein-glyceraldehyde 3-phosphate dehydrogenase
Protein
Compartment
(1-Mf)
t1/2, s
D,μm2/s
EGFP-GAPDH-WT
cyto
0.18 ± 0.038
0.19 ± 0.055
27.17 ± 1.632
EGFP-GAPDH-Y94A
cyto
0.17 ± 0.129
0.09 ± 0.014
43.76 ± 7.158
EGFP-GAPDH-S98A
cyto
0.20 ± 0.009
0.12 ± 0.016
29.15 ± 3.885
EGFP-GAPDH-T99A
cyto
0.11 ± 0.062
0.11 ± 0.023
35.52 ± 8.133
EGFP-GAPDH-T99I
cyto
0.17 ± 0.06
0.22 ± 0.041
17.46 ± 1.733
EGFP-GAPDH-WT
nuclei
0.62 ± 0.048
1.41 ± 0.033
2.91 ± 0.331
EGFP-GAPDH-Y94A
nuclei
0.23 ± 0.127
0.09 ± 0.034
39.05 ± 6.291
EGFP-GAPDH-S98A
nuclei
0.35 ± 0.136
0.13 ± 0.018
26.57 ± 3.747
EGFP-GAPDH-T99A
nuclei
0.38 ± 0.120
0.11 ± 0.027
33.01 ± 9.209
EGFP-GAPDH-T99I
nuclei
0.29 ± 0.028
0.21 ± 0.059
14.97 ± 1.855
Table 4 Average potential energy, kinetic energy, total energy and temperature calculated from dynamic simulations
Polypeptide variant
Average potential (kcal)
Average kinetic energy (kcal)
Average total energy (kcal)
Average temperature (°C)
T99
-5021.8
97.1
-4924.7
293.4
A99
-5007.3
93.7
-4913.6
293.7
I99
-4997.8
101.4
-4896.4
293.3
Citation: Phadke M, Krynetskaia N, Mishra A, Barrero C, Merali S, Gothe SA, Krynetskiy E. Disruption of NAD+ binding site in glyceraldehyde 3-phosphate dehydrogenase affects its intranuclear interactions. World J Biol Chem 2015; 6(4): 366-378