Glucosamine 6-phosphate deaminase in normal human erythrocytes.
In the course of an investigation of hexosamine catabolism in the human malaria parasite, Plasmodium falciparum, it became apparent that a basic understanding of the relevant enzymatic reactions in the host erythrocyte is lacking. To acquire the necessary basic knowledge, we have determined the activities of several enzymes involved in hexosamine metabolism in normal human red blood cells. In the present communication we report the results of studies of glucosamine 6-phosphate deaminase (GlcN6-P) using a newly developed sensitive radiometric assay. The mean specific activity in extracts of fresh erythrocytes assayed within 4h of collection was 14.7 nmol/h/mg protein, whereas preparations from older erythrocytes that had been stored at 4 degrees C for up to 4 weeks had a mean specific activity of 6.2 nmol/h/mg. Characterization of the deaminase by chromatofocusing gave a pI of 8.55. The enzyme was optimally active at pH 9.0 and had a Km of 41 microM. The metal chelators EDTA and EGTA were non-inhibitory; however, inhibition was observed in the presence of metal ions, especially Cu2+, Ni2+ and Zn2+. In addition, the deaminase was also inhibited by several sugar phosphates including the reaction product, fructose 6-phosphate.
Weidanz
J A
JA
Department of Microbiology, School of Medicine, University of Alabama at Birmingham 35294-2170, USA.
Campbell
P
P
DeLucas
L J
LJ
Jin
J
J
Moore
D
D
Rodén
L
L
Yu
H
H
Heilmann
E
E
Vezza
A C
AC
D019747:Aldose-Ketose Isomerases; D002238:Carbohydrate Epimerases / Q000097:blood* / Q000187:drug effects; D002241:Carbohydrates / Q000494:pharmacology; D002469:Cell Separation; D002852:Chromatography, Ion Exchange; D004912:Erythrocytes / Q000201:enzymology*; D005434:Flow Cytometry; D006801:Humans; D007700:Kinetics; D016778:Malaria, Falciparum / Q000201:enzymology; D008670:Metals / Q000494:pharmacology; D013696:Temperature
D002241:Carbohydrates; D008670:Metals; C020582:glucosamine-6-phosphate isomerase; D002238:Carbohydrate Epimerases; D019747:Aldose-Ketose Isomerases