Written in English
|Statement||by Jean Dolde Jenkins.|
|LC Classifications||Microfilm 84/112 (Q)|
|The Physical Object|
|Pagination||viii, 120 leaves|
|Number of Pages||120|
|LC Control Number||84151374|
Association of Phosphofructokinase and Aldolase with the Membrane of the Intact Erythrocyte* (Received for publication, Janu) Jean D. Jenkins$, Donald P. Madden, and Theodore L. Steckg. The interaction of muscle phosphofructokinase with human erythrocyte ghost membranes was investigated. Scatchard analysis of the binding reveals only a single class of binding sites numbering approximately 4 x 10 5 sites/ghost with an association constant of 2 x 10 7 MMaximum binding is observed below pH and is complete within 30 min at 4°, 24°, and 37°C; binding is inhibited by high Cited by: Association of phosphofructokinase and aldolase with the membrane of the intact erythrocyte. Jenkins JD, Madden DP, Steck TL. J Biol Chem, (15), 01 Aug Cited by 33 articles | PMID: Cited by: phosphate aldolase (4-ll), and phosphoglycerate kinase (12) have been shown to be bound to the human erythrocyte membrane. The binding of these enzymes is tight, but revers- ible, sensitive to high ionic strength, and pH-dependent. Cer- tain physiologically significant metabolites influence the bind-.
A study has been carried out on the association of aldolase with the human erythrocyte membrane. It has been shown that the conditions employed during hypotonic hemolysis affect the amount of aldolase that remains bound to the cell membrane. Thus, the in vivo nature of this binding cannot be ascertained by this technique. Therefore, a method has been developed in which aldolase is crosslinked with glutaraldehyde . of the human erythrocyte. This ,dalton glycoprotein mediates the exchange of anions across the membrane (see Refs. for review). Furthermore, in vitro, the cytoplasmic domain of band 3 is the exclusive site of binding of aldolase (6, 71, glyceraldehydeP dehydrogenase (81, and probably phosphofructokinase (9) and hemoglobin (). Observations that aldolase and phosphofructokinase (PFK) also associate with the same NH 2-terminal sequence of band 3 then prompted interest in the possible assembly of a GE complex on the cytoplasmic domain of band 3, and this interest subsequently led to investigations into whether the above enzymes are, in fact, membrane-bound in the intact cell. As noted previously, the association of aldolase with the cytoplasmic domain of band 3 resulted in the reversible inhibition of aldolase identify whether the recombinant CDB3-(His) 6 has the same function to inhibit the aldolase activity as nature CDB3 in erythrocyte, the aldolase activity was assayed after association with CDB3-(His) 6.
J.D. Jenkis, D.P. Madden and T.L. Steck, Association of phosphofructokinase and aldolase with the membrane of the intact erythrocyte, J. Biol. Chem. (). Google Scholar 6. Observations that aldolase and phosphofructokinase (PFK) also associate with the same NH 2-terminal sequence of band 3 then prompted interest in the possible assembly of a GE complex on the cytoplasmic domain of band 3, and this interest subsequently led to investigations into whether the above enzymes are, in fact, membrane-bound in the. human erythrocytes, freshly drawn blood was fixed and stained with Abs to GAPDH, aldolase, phosphofructokinase (PFK), pyruvate kinase (PK), lactate dehydrogenase (LDH), carbonic anhydr Hb, and band 3 (AEl). Confocal microscopy revealed that in cells where band 3 displays its expected membrane staining and Hb is. We wish to elaborate a novel mechanism of metabolic regulation mediated by cytoplasmic tyrosine phosphatases and kinases. Briefly we propose that phosphofructokinase, aldolase, and glyceraldehydephosphate dehydrogenase (G3PDH) bind reversibly to the N-terminus of the cytoplasmic domain of band 3. Once the enzymes are bound, they are inhibited; however, upon .