This chapter describes the purification and the properties of hexokinase found in rabbit reticulocytes. This enzyme shows a marked decrease during cell aging, and two distinct molecular forms (hexokinase Ia and Ib) are found in reticulocytes, but only one of these is consistently present in the mature rabbit red cell. With glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in excess, the rate of glucose phosphorylation is equivalent to the rate of NADP+ reduction, which is measured by the absorbance increase at 340 nm. For each molecule of glucose utilized, two molecules of NADP+ are reduced. Purification procedure for hexokinase from rabbit red blood cells involves preparation of hemolysate, batch treatment with DEAE-Sephadex A-50, ammonium sulfate fractionation, affinity chromatography on Sepharose-N-amino-hexanoylglucosamine column, and preparative polyacrylamide gel electrophoresis. Both hexokinase Ia and Ib can be stabilized by glucose, fructose, glycerol, and sulfhydryl protecting agents, as can hexokinase from mature rabbit red cells. Series: Methods in Enzymology (Book 90) Hardcover: 640 pages Publisher: Academic Press; Language: English ISBN-10: 0121819906 ISBN-13: 978-0121819903 Product Dimensions: 6 x 1.4 x 9 inches Link Download http://nitroflare.com/view/998EB032602E115https://drive.google.com/drive/folders/1yLBzZ1rSQoNjmWeJTZ3WGQHg04L1
