Literature summary for 1.2.1.12 extracted from

Baker, B.Y.; Shi, W.; Wang, B.; Palczewski, K.
High-resolution crystal structures of the photoreceptor glyceraldehyde 3-phosphate dehydrogenase (GAPDH) with three and four-bound NAD molecules (2014), Protein Sci., 23, 1629-1639 .

Application

Application	Comment	Organism
drug development	GAPDH is an important drug target	Bos taurus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified native tetrameric enzyme with either three or four bound NAD+ molecules, bGAPDH can be crystallized directly from isotonic extracts of ROS, X-ray diffraction structure determination and analysis at 1.93 A and 1.54 A resolution, respectively, molecular replacement using the coordinates of one monomer of rabbit-muscle GAPDH, PDB ID 1J0X, without ligand and water to search the initial model, modelling	Bos taurus

Crystallization (Comment)

Organism

purified native tetrameric enzyme with either three or four bound NAD+ molecules, bGAPDH can be crystallized directly from isotonic extracts of ROS, X-ray diffraction structure determination and analysis at 1.93 A and 1.54 A resolution, respectively, molecular replacement using the coordinates of one monomer of rabbit-muscle GAPDH, PDB ID 1J0X, without ligand and water to search the initial model, modelling

Bos taurus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.153	-	D-glyceraldehyde 3-phosphate	pH and temperature not specified in the publication	Bos taurus
0.255	-	NAD+	pH and temperature not specified in the publication	Bos taurus

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
150000	-	gel filtration	Bos taurus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
D-glyceraldehyde 3-phosphate + phosphate + NAD+	Bos taurus	-	3-phospho-D-glyceroyl phosphate + NADH + H+	-	?

Organism

Organism	UniProt	Comment	Textmining
Bos taurus	Q2KJE5	-	-

Purification (Commentary)

Purification (Comment)	Organism
native enzyme from bovine retinas using NAD+-agarose affinity chromatography	Bos taurus

Source Tissue

Source Tissue	Comment	Organism	Textmining
eye	-	Bos taurus	-
photoreceptor	-	Bos taurus	-
retina	-	Bos taurus	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-glyceraldehyde 3-phosphate + phosphate + NAD+	-	Bos taurus	3-phospho-D-glyceroyl phosphate + NADH + H+	-	?

Subunits

Subunits	Comment	Organism
homotetramer	4 * 36000, SDS-PAGE	Bos taurus
More	structure analysis, overview. Dual side-chain conformations are observed in Ser207 in subunits of O, Q, and R of the bGAPDH(NAD)3	Bos taurus

Synonyms

Synonyms	Comment	Organism
GAPDH	-	Bos taurus
glyceraldehyde-3-phosphate dehydrogenase	-	Bos taurus

Cofactor

Cofactor	Comment	Organism	Structure
NAD+	differing occupancy by NAD, bGAPDH(NAD)4, and bGAPDH(NAD)3 in the homotetramer, structure analysis, overview. Importance of Phe34 in NAD+ binding, Phe34 is stabilized in the presence of NAD+ but displays greater mobility in its absence. The oxidative state of the active site Cys149 residue is regulated by NAD+ binding, because this residue is found oxidized in the absence of dinucleotide. The distance between Cys149 and His176 decreases upon NAD binding and Cys149 remains in a reduced state when NAD+ is bound. Dual side-chain conformations are observed in Ser207 in subunits of O, Q, and R of the bGAPDH(NAD)3. Residues Pro33 and Phe34 form a bottleneck for NAD+ binding	Bos taurus

General Information

General Information	Comment	Organism
malfunction	substitution of Phe34 with smaller side chain (e.g. Gly or Leu), or polar residue (e.g. Thr) abolishes the NAD+ binding affinity, or reduce the protein's catalytic efficiency	Bos taurus
additional information	importance of Phe34 in NAD+ binding, Phe34 is stabilized in the presence of NAD+ but displays greater mobility in its absence. The oxidative state of the active site Cys149 residue is regulated by NAD+ binding, because this residue is found oxidized in the absence of dinucleotide. The distance between Cys149 and His176 decreases upon NAD binding and Cys149 remains in a reduced state when NAD+ is bound, cofactor binding and active site structures, catalytic mechanism, overview	Bos taurus
physiological function	the enzyme plays a central role in glycolysis, and nonglycolytic processes such as nuclear RNA transport, DNA replication/repair, membrane fusion and cellular apoptosis	Bos taurus