The Antarctic Psychrobacter sp. TAD1 has two cold-active glutamate dehydrogenases with different cofactor specificities. Characterisation of the NAD+-dependent enzyme.
Psychrobacter sp. TAD1 is a psychrotolerant bacterium from Antarctic frozen continental water that grows from 2 to 25 degrees C with optimal growth rate at 20 degrees C. The new isolate contains two glutamate dehydrogenases (GDH), differing in their cofactor specificities, subunit sizes and arrangements, and thermal properties. NADP+-dependent GDH is a hexamer of 47 kDa subunits and it is comparable to other hexameric GDHs of family-I from bacteria and lower eukaria. The NAD+-dependent enzyme, described in this communication, has a subunit weight of 160 kDa and belongs to the novel class of GDHs with large size subunits. The enzyme is a dimer; this oligomeric arrangement has not been reported previously for GDH. Both enzymes have an apparent optimum temperature for activity of approximately 20 degrees C, but their cold activities and thermal labilities are different. The NAD+-dependent enzyme is more cold active: at 10 C it retains 50% of its maximal activity, compared with 10% for the NADP+-dependent enzyme. The NADP+-dependent enzyme is more heat stable, losing only 10% activity after heating for 30 min, compared with 95% for the NAD+-dependent enzyme. It is concluded that in Psychrobacter sp. TAD1 not only does NAD+-dependent GDH have a novel subunit molecular weight and arrangement, but that its polypeptide chains are folded differently from those of NADP+-dependent GDH, providing different cold-active properties to the two enzymes.
Camardella
Laura
L
Institute of Protein Biochemistry and Enzymology, C.N.R. Via Marconi 12, I-80125 Naples, Italy.
Di Fraia
Raffaela
R
Antignani
Antonella
A
Ciardiello
M Antonietta
MA
di Prisco
Guido
G
Coleman
Julie K
JK
Buchon
Laurent
L
Guespin
Janine
J
Russell
Nicholas J
NJ
D000222:Adaptation, Physiological / Q000502:physiology; D000595:Amino Acid Sequence; D000864:Antarctic Regions; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D003080:Cold Temperature; D020563:Gammaproteobacteria / Q000201:enzymology*; D005969:Glutamate Dehydrogenase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D018086:Glutamate Dehydrogenase (NADP+) / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism*; D009249:NADP / Q000378:metabolism*
D001426:Bacterial Proteins; D009243:NAD; D009249:NADP; D005969:Glutamate Dehydrogenase; D018086:Glutamate Dehydrogenase (NADP+)