Characterization of glycolytic enzyme interactions with murine erythrocyte membranes in wild-type and membrane protein knockout mice. Blood 2008 Nov 01;112(9):3900-6
Date
08/14/2008Pubmed ID
18698006Pubmed Central ID
PMC2572807DOI
10.1182/blood-2008-03-146159Scopus ID
2-s2.0-55749086024 (requires institutional sign-in at Scopus site) 93 CitationsAbstract
Previous research has shown that glycolytic enzymes (GEs) exist as multienzyme complexes on the inner surface of human erythrocyte membranes. Because GE binding sites have been mapped to sequences on the membrane protein, band 3, that are not conserved in other mammalian homologs, the question arose whether GEs can organize into complexes on other mammalian erythrocyte membranes. To address this, murine erythrocytes were stained with antibodies to glyceraldehyde-3-phosphate dehydrogenase, aldolase, phosphofructokinase, lactate dehydrogenase, and pyruvate kinase and analyzed by confocal microscopy. GEs were found to localize to the membrane in oxygenated erythrocytes but redistributed to the cytoplasm upon deoxygenation, as seen in human erythrocytes. To identify membrane proteins involved in GE assembly, erythrocytes from mice lacking each of the major erythrocyte membrane proteins were examined for GE localization. GEs from band 3 knockout mice were not membrane associated but distributed throughout the cytoplasm, regardless of erythrocyte oxygenation state. In contrast, erythrocytes from mice lacking alpha-spectrin, ankyrin, protein 4.2, protein 4.1, beta-adducin, or dematin headpiece exhibited GEs bound to the membrane. These data suggest that oxygenation-dependent assembly of GEs on the membrane could be a general phenomenon of mammalian erythrocytes and that stability of these interactions depends primarily on band 3.
Author List
Campanella ME, Chu H, Wandersee NJ, Peters LL, Mohandas N, Gilligan DM, Low PSMESH terms used to index this publication - Major topics in bold
Amino Acid SequenceAnimals
Anion Exchange Protein 1, Erythrocyte
Base Sequence
DNA, Complementary
Erythrocyte Membrane
Fructose-Bisphosphate Aldolase
Glyceraldehyde-3-Phosphate Dehydrogenases
Glycolysis
L-Lactate Dehydrogenase
Mice
Mice, Knockout
Molecular Sequence Data
Multienzyme Complexes
Oxygen
Phosphofructokinases
Pyruvate Kinase
Sequence Homology, Amino Acid
