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1868   Part XII  Hemostasis and Thrombosis


        molecule” that collects binding partners and localizes them adjacent   the affinity of spectrin for adducin-actin complexes is greater than for
        to the plasma membrane. Partners bound by filamin members include   either  actin  or  adducin  alone.  Platelet  glycoproteins  involved  in
        the small GTPases, RalA, Rac, Rho, and Cdc42, with RalA binding   attaching spectrin to the membrane remain to be defined.
        in  a  GTP-dependent  manner;  the  exchange  factors Trio  and Toll;
        kinases such as PAK1; phosphatases; and transmembrane proteins.
        Most partner proteins are bound within the C-terminus portion of
        filamin.                                              REFERENCES
           Central to the structural organization of the resting platelet is an
        interaction between filamin and the cytoplasmic tail of the GPIbα   1.  Bluteau D, Lordier L, Di Stefano A, et al: Regulation of megakaryocyte
                                  27
        subunit of the GPIb-IX-V complex.  The second rod domain (repeat   maturation and platelet formation. J Thromb Haemost 7:227, 2009.
        17) of filamin has a binding site for the cytoplasmic tail of GPIbα.   2.  Schulze H, Korpal M, Hurov J, et al: Characterization of the megakaryo-
        The interaction between filamin A and GPIbα occurs at the atomic   cyte demarcation membrane system and its role in thrombopoiesis. Blood
        level. Repeat 17 of filamin A has a groove between certain β-sheet   107:3868, 2006.
        strands  that  forms  a  pocket  for  the  GPIbα  tail  (see  Fig.  124.10).   3.  Lu SJ, Li F, Yin H, et al: Platelets generated from human embryonic stem
        Binding between filamin A and GPIbα is driven by entropic forces,   cells are functional in vitro and in the microcirculation of living mice.
        and the alignment and specificity are provided by large residues that   Cell Res 2011.
        create  a  lock-and-key  fit  between  the  two  molecules. Whereas  the   4.  Takayama N, Nishimura S, Nakamura S, et al: Transient activation of
        interaction between one filamin A subunit and GPIbα has an affinity   c-MYC expression is critical for efficient platelet generation from human
        of approximately 10 µM, high-affinity binding (10 nM) occurs when   induced pluripotent stem cells. J Exp Med 207:2817, 2010.
        each filamin A subunit in a molecule and both GPIbα chains in a   5.  Italiano  JE,  Jr,  Lecine  P,  Shivdasani  RA,  et al:  Blood  platelets  are
        vWF  receptor  are  engaged.  Studies  have  shown  that  the  bulk  of   assembled principally at the ends of proplatelet processes produced by
        platelet filamin (>90%) is in complex with GPIbα. This interaction   differentiated megakaryocytes. J Cell Biol 147:1299, 1999.
        has  three  consequences.  First,  it  positions  filamin’s  self-association   6.  Schwer H, Lecine P, Tiwari S, et al: A lineage-restricted and divergent b
        domain  and  associated  partner  proteins  at  the  plasma  membrane   tubulin isoform is essential for the biogenesis, structure and function of
        while  dangling  filamin’s  actin  binding  sites  into  the  cytoplasm.   mammalian blood platelets. Curr Biol 11:579, 2001.
        Second, because a large fraction of filamin is bound to actin, it aligns   7.  Patel S, Richardson J, Schulze H, et al: Differential roles of microtubule
        the  GPIb-IX-V  complexes  on  the  surface  of  the  platelet  over  the   assembly and sliding in proplatelet formation by megakaryocytes. Blood
        underlying filaments (see Fig. 124.9B). Third, because the filamin   106:4076, 2005.
        linkages between actin filaments and the GPIb-IX-V complex pass   8.  Patel S, Hartwig J, Italiano J, Jr: The biogenesis of platelets from mega-
        through the pores of the spectrin lattice, it restrains the molecular   karyocyte proplatelets. J Clin Invest 115:3348, 2006.
        movement of the spectrin strands in this lattice and holds the lattice   9.  Schulze H, Dose M, Korpal M, et al: RanBP10 is a cytoplasmic guanine
        in compression. The filamin-GPIbα connection is essential for the   nucleotide exchange factor that modulates noncentrosomal microtubules.
        formation and release of discoid platelets by megakaryocytes; platelets   J Biol Chem 283:14109, 2008.
        lacking this connection are large and fragile and are produced in low   10.  Chen Z, Naveiras O, Balduini A, et al: The May-Hegglin anomaly gene
        numbers. However, the role of the filamin-vWF receptor connection   MYH9 is a negative regulator of platelet biogenesis modulated by the
        in  platelet  construction  is  unknown.  Because  a  low  number  of   Rho-ROCK pathway. Blood 110:171, 2007.
        Bernard-Soulier platelets form and release from megakaryocytes, it   11.  Eckly A, Strassel C, Freund M, et al: Abnormal megakaryocyte morphol-
        can be argued that this connection is a late event in the maturation   ogy  and  proplatelet  formation  in  mice  with  megakaryocyte-restricted
        process and is not required for platelet shedding. Both filamin and   MYH9 inactivation. Blood 113:3182, 2009.
        GPIbα are synthesized early, but linkage between the two may not   12.  Leon  C,  Eckly  A,  Hechler  B,  et al:  Megakaryocyte-restricted  MYH9
        occur until later, perhaps as late as the final stages of platelet shedding.   inactivation  dramatically  affects  hemostasis  while  preserving  platelet
        Bernard-Soulier patients have an autosomal recessive bleeding disor-  aggregation and secretion. Blood 110:3183, 2007.
        der characterized by low platelet count with giant platelets, under-  13.  Chen Z, Shivdasani RA: Regulation of platelet biogenesis: insights from
        scoring the importance of the filamin-GPIbα connection in stabilizing   the May-Hegglin anomaly and other MYH9-related disorders. J Thromb
        the membrane skeleton in platelet biology.               Haemost 7:272, 2009.
           Aside  from  the  erythrocyte,  the  platelet  is  the  only  cell  whose   14.  Richardson  J,  Shivdasani  R,  Boers  C,  et al:  Mechanisms  of  organelle
        membrane skeleton has been visualized at high resolution. Like the   transport  and  capture  along  proplatelets  during  platelet  production.
        erythrocyte,  the  platelet  membrane  skeleton  is  a  self-assembly  of   Blood 106:4066, 2005.
        elongated spectrin strands (see Fig. 124.9) that interconnect through   15.  Patel-Hett S, Wang H, Begonja AJ, et al: The spectrin-based membrane
        binding  to  actin  filaments.  Platelets  express  approximately  2000   skeleton  stabilizes  mouse  megakaryocyte  membrane  systems  and  is
        spectrin molecules. Although considerably less is known about how   essential for proplatelet and platelet formation. Blood 2011.
        the  spectrin-actin  network  forms  and  is  connected  to  the  plasma   16.  Junt T, Schulze H, Chen Z, et al: Dynamic visualization of thrombopoi-
        membrane in the platelet relative to the erythrocyte, certain differ-  esis within bone marrow. Science 317:1767, 2007.
        ences  between  the  two  membrane  skeletons  have  been  identified.   17.  Thon  JN,  Montalvo  A,  Patel-Hett  S,  et al:  Cytoskeletal  mechanics
        First, the spectrin strands composing the platelet membrane skeleton   of  proplatelet  maturation  and  platelet  release.  J  Cell  Biol  191:861,
        interconnect using the ends of long actin filaments instead of short   2010.
        actin oligomers. These ends arrive at the plasma membrane originat-  18.  Schwertz  H,  Koster  S,  Kahr  WH,  et al:  Anucleate  platelets  generate
        ing  from  filaments  in  the  cytoplasm.  Hence  the  spectrin  lattice  is   progeny. Blood 115:3801, 2010.
        assembled into a continuous network by its association with actin   19.  Larson MK, Watson SP: A product of their environment: do megakaryo-
        filaments.  Second,  tropomodulins  are  not  expressed  at  sufficiently   cytes rely on extracellular cues for proplatelet formation? Platelets 17:435,
        high levels, if at all, to have a major role in capping the pointed ends   2006.
        of the platelet actin filaments. Instead, biochemical experiments have   20.  Larson MK, Watson SP: Regulation of proplatelet formation and platelet
        revealed that a substantial number (≈2000) of these ends are free in   release by integrin alpha IIb beta3. Blood 108:1509, 2006.
        the resting platelet. Third, although little tropomodulin protein is   21.  Goldenson B, Kirsammer G, Stankiewicz MJ, et al: Aurora kinase A is
        expressed, α adducin and γ adducin are abundantly expressed and   required for hematopoiesis, but is dispensable for murine megakaryocyte
        appear to cap many of the barbed ends of the filaments composing   endomitosis and differentiation. Blood 125:2141, 2015.
        the resting actin cytoskeleton. Adducin is a key component of the   22.  Dore LC, Crispino JD: Transcription factor networks in erythroid cell
        membrane skeleton, forming a triad complex with spectrin and actin.   and megakaryocyte development. Blood 2011.
        Capping of barbed filament ends by adducin also serves the function   23.  Hartwig JH: The platelet: form and function. Semin Hematol 43:S94,
        of targeting them to the spectrin-based membrane skeleton because   2006.
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