1-13-10 RBC membrane, cytoskeleton and physiology 1.

Describe how abnormal production of metabolites of glycolysis (ATP, NADH, NADPH due to genetic or en!ironmental stress, in "#$ affect ion (Na, %, and $a transport, o&ygen affinity and deformability (fle&ibility , leading to hemolytic anemia. Glucose is the major fuel of the rbc !t enters the cell by facilitated diffusion and does not require insulin for uptake "0-"#$ of this glucose is metaboli%ed to lactate by the glycolytic path&ay' the rest is taken up by the he(os monophosphate )pentose* path&ay to produce +,-./ ,s mentioned before, the three metabolites of glycolysis are ,0., +,-/, and +,-./ )there1s also 2,3-B.G, &hich maintains o(ygen affinity and is not important here* /ere1s &hy each are important for proper rbc function and morphology3 ATP3 t&o moles of ,0. are needed to put one glucose through glycolysis !t is used up by Na/K pumps )to maintain high intracellular 4 and lo& intracellular +a* as &ell as CaATPase pumps )to maintain lo& intracellular Ca* Reduced Ca pump function is characteristic of sic'le cell anemia Conse5uently a deficiency of ,0. &ill cause' ,ccumulation of Ca inside rbc1s-> increases membrane-associated calcium>increased membrane rigidity->reduced deformability->hemolysis during rbc transit through small capillaries ,ccumulation of Ca inside rbc1s -> (ardos effect )intracellular Ca causes opening of 4 channels, resulting in an efflux of K from the cell* -> high intracellular Ca and lo& intracellular 4 -> dehydrated cell Pyru!ate 'inase deficiency is the most common enzyme abnormality in glycolysis !t results in lo production of ATP, &hich means rbc1s easily hemoly%e Conse5uently, patients &ith .4 deficiency ha6e lifelong hemolytic anemia NADH3 functions to con6ert methemoglobin )&ith 7e38* to hemoglobin )7e28* 6ia +,-/-cytochrome b# reductase -eficiency in +,-/ or that en%yme &ill cause' ,ccumulation of methemoglobin that can1t bind o(ygen->cyanosis 9pecifically, a deficiency in +,-/-cytochrome b# reductase>methemoglobinemia in response to o(idant drugs such i e malarial prophylactic agents prima5uine and chloro5uine NADPH3 technically, +,-./ is not produced 6ia glycolysis, but 6ia the he(ose monophosphate )pentose* phosphate path&ay 0he function of +,-./ is to regernerate !"# from !""! in a reaction cataly%ed by glutathione reductase :oreo6er, +,-./ detoxes $ydrogen peroxide during infection !t is made from G-;-. 6ia G-;-. dehydrogenase , deficiency in +,-./ results from G-;-. deficiency and &ould therefore cause' 'hemolytic anemia )see ne(t objecti6e* ). Describe how o&idant drugs or infection causes hemolysis of rbc in patients with deficiency of (*+*P dehydrogenase. (*+*P dehydrogenase deficiency leads to a deficiency in NA%P# 9ince +,-./ functions to con6ert G99G )o(idi%ed glutathione* to G9/ )reduced glutathione, &hich protects rbc1s against

o(idant injury by R<9*, the result is chronic hemolytic anemia !mportant points about this condition3 !t is the most common cause of $emolytic anemia !t is =-linked inherited .atients %& N&T "#&' C()N)CA( manifestations of $emolytic anemia unless t$ey are exposed to infection or are taking oxidant drugs 0hus, G-;-. dehydrogenase deficiency is characteri%ed as >hemolytic anemia due to inability to deto(ify o(idi%ing agents ? -uring infection, macrophages generate pero(ide that diffuses into rbc1s, resulting in crosslinking of membrane proteins and lipid pero(idation 0his is &hat G9/ pre6ents 0he positi6e effect of G-;-. dehydrogenase deficiency is that female carriers are resistant to malaria* ,. Discuss the role of flipase (aminophospholipid translocase in asymmetric distribution of phospholipids in the bilayer of rbc membrane. -lipase is an ,0.-dependent en%yme in charge of mo+ing bot$ p$osp$atidyl et$anolamine ,PEand p$osp$atidyl serine ,PS- to t$e internal leaflet of t$e bilayer. 0his is the reason &hy .@ and .9 are only present on the inner leaflet )and phosphatidylcholine ).C* and sphingomyelin )9:* are on the outer leaflet* .. Discuss how abnormal presence of P/ (phosphatidyl serine in the e&ternal leaflet of the bilayer of rbc initiates coagulation. ,s mentioned abo6e, .9 )and .@* are only supposed to be found on the inner leaflet 0hat1s because .9 on the e(ternal leaflet can initiate a cascade of reactions in6ol6ed in coagulation 0his is seen in sickle cell disease, &here a small amount of .9 in the e(ternal leaflet leads to coagulation->clot formation->6asoocclusion 0. "ecogni1e names of cytos'eletal proteins (spectrin, an'yrin, and actin and their role in maintaining shape and deformability of rbc2s. Remember the main cytoskeletal proteins3 spectrin, an'yrin, actin and protein ..1 0he cytoskeleton helps maintain s$ape and deformability of the rbc by interacting &ith integral proteins Band 3 and glycophorin ,ny changes to these cytoskeletal proteins increases susceptibility to hemolysis 7or e(ample, see objecti6e ; +. Discuss how genetic defect in cytos'eletal proteins (e.g. spectrin leads to hemolytic anemia. Reduced content of spectrin->hereditary spherocytosis )autosomal dominant condition resulting in spherocytes causing mild anemia* /pectrin molecule unable to form tetramer resulting in &eakened cytoskeleton->hereditary elliptocytosis )autosomal dominant condition resulting in elliptocytes &ith mild anemia*

3. Discuss how abnormal accumulation of cholesterol in "#$ membrane, due to alcoholism, leads to hemolytic anemia. !ncrease in cholesterol and sphingomyelin->acanthocytosis )autosomal recessi6e condition resulting in rbc1s &ith thorny projections called spiculesA occurs &ith ,BC</<B!C li6er disease* 4. Discuss how single point mutation in the beta*chain of hemoglobin in sic'le blood cells leads to hemolytic anemia, !aso*occlusion and disease pathophysiology. , mutation of the /-globin c$ain at t$e 0t$ AA glu ,polar-->+al ,nonpolar- produces Hb/ /b9 tends to occur at lo& o(ygen tension, resulting in the formation of long insoluble fibers that change the shape and reduce the deformability of sickled cells 0his leads to occlusion of capillaries )caused by adhesion of cell integrins to CC,:-11s on endothelial cells* and hemolysis 0he occlusions can cause acute pain crises, acute chest syndromes, and stroke !n summary, the t&o main clinical features of sickle cell disease )9C-* are c$ronic $emolytic anemia and +asoocclusi+e pain crises T$ere is no specific treatment for "C% 0reatments for 9C- include reducing fe6er &ith analgesics, correcting acidosis )lo& p/ promotes polymeri%ation of /b9*, increasing fluid intake )slo&s do&n polymeri%ation*, treatment &ith hydro(yurea )increases amount of fetal hemoglobin*, pre6enting adhesion of rbc1s to 6ascular endothelium, leukotriene antagonists to lo&er asthma and reacti6e air&ay disease, and gene therapy