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. 1990 Sep 1;111(3):1089–1105. doi: 10.1083/jcb.111.3.1089

Human endothelial actin-binding protein (ABP-280, nonmuscle filamin): a molecular leaf spring

PMCID: PMC2116286  PMID: 2391361

Abstract

Actin-binding protein (ABP-280, nonmuscle filamin) is a ubiquitous dimeric actin cross-linking phosphoprotein of peripheral cytoplasm, where it promotes orthogonal branching of actin filaments and links actin filaments to membrane glycoproteins. The complete nucleotide sequence of human endothelial cell ABP cDNA predicts a polypeptide subunit chain of 2,647 amino acids, corresponding to 280 kD, also the mass derived from physical measurements of the native protein. The actin-binding domain is near the amino-terminus of the subunit where the amino acid sequence is similar to other actin filament binding proteins, including alpha-actinin, beta-spectrin, dystrophin, and Dictyostelium abp-120. The remaining 90% of the sequence comprises 24 repeats, each approximately 96 residues long, predicted to have stretches of beta-sheet secondary structure interspersed with turns. The first 15 repeats may have substantial intrachain hydrophobic interactions and overlap in a staggered fashion to yield a backbone with mechanical resilience. Sequence insertions immediately before repeats 16 and 24 predict two hinges in the molecule near points where rotary-shadowed molecules appear to swivel in electron micrographs. Both putative hinge regions are susceptible to cleavage by proteases and the second also contains the site that binds the platelet glycoprotein Ib/IX complex. Phosphorylation consensus sequences are also located in the hinges or near them. Degeneracy within every even- numbered repeat between 16 and 24 and the insertion before repeat 24 may convert interactions within chains to interactions between chains to account for dimer formation within a domain of 7 kD at the carboxy- terminus. The structure of ABP dimers resembles a leaf spring. Interchain interactions hold the leaves firmly together at one end, whereas intrachain hydrophobic bonds reinforce the arms of the spring where the leaves diverge, making it sufficiently stiff to promote high- angle branching of actin filaments. The large size of the leaves, their interruption by two hinges and flexible actin-binding site, facilitate cross-linking of widely dispersed actin filaments.

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Selected References

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  1. Baron M. D., Davison M. D., Jones P., Critchley D. R. The sequence of chick alpha-actinin reveals homologies to spectrin and calmodulin. J Biol Chem. 1987 Dec 25;262(36):17623–17629. [PubMed] [Google Scholar]
  2. Blackshear P. J., Nairn A. C., Kuo J. F. Protein kinases 1988: a current perspective. FASEB J. 1988 Nov;2(14):2957–2969. doi: 10.1096/fasebj.2.14.2972578. [DOI] [PubMed] [Google Scholar]
  3. Boxer L. A., Richardson S., Floyd A. Identification of actin-binding protein in membrane of polymorphonuclear leukocytes. Nature. 1976 Sep 16;263(5574):249–251. doi: 10.1038/263249a0. [DOI] [PubMed] [Google Scholar]
  4. Brotschi E. A., Hartwig J. H., Stossel T. P. The gelation of actin by actin-binding protein. J Biol Chem. 1978 Dec 25;253(24):8988–8993. [PubMed] [Google Scholar]
  5. Byers T. J., Husain-Chishti A., Dubreuil R. R., Branton D., Goldstein L. S. Sequence similarity of the amino-terminal domain of Drosophila beta spectrin to alpha actinin and dystrophin. J Cell Biol. 1989 Oct;109(4 Pt 1):1633–1641. doi: 10.1083/jcb.109.4.1633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carroll R. C., Gerrard J. M. Phosphorylation of platelet actin-binding protein during platelet activation. Blood. 1982 Mar;59(3):466–471. [PubMed] [Google Scholar]
  7. Chen M., Stracher A. In situ phosphorylation of platelet actin-binding protein by cAMP-dependent protein kinase stabilizes it against proteolysis by calpain. J Biol Chem. 1989 Aug 25;264(24):14282–14289. [PubMed] [Google Scholar]
  8. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  9. Chothia C. Coiling of beta-pleated sheets. J Mol Biol. 1983 Jan 5;163(1):107–117. doi: 10.1016/0022-2836(83)90031-1. [DOI] [PubMed] [Google Scholar]
  10. Chou P. Y., Fasman G. D. Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol Relat Areas Mol Biol. 1978;47:45–148. doi: 10.1002/9780470122921.ch2. [DOI] [PubMed] [Google Scholar]
  11. Condeelis J., Vahey M., Carboni J. M., DeMey J., Ogihara S. Properties of the 120,000- and 95,000-dalton actin-binding proteins from Dictyostelium discoideum and their possible functions in assembling the cytoplasmic matrix. J Cell Biol. 1984 Jul;99(1 Pt 2):119s–126s. doi: 10.1083/jcb.99.1.119s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cortese J. D., Frieden C. Microheterogeneity of actin gels formed under controlled linear shear. J Cell Biol. 1988 Oct;107(4):1477–1487. doi: 10.1083/jcb.107.4.1477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Corwin H. L., Hartwig J. H. Isolation of actin-binding protein and villin from toad oocytes. Dev Biol. 1983 Sep;99(1):61–74. doi: 10.1016/0012-1606(83)90254-3. [DOI] [PubMed] [Google Scholar]
  14. Dorfman D. M., Zon L. I., Orkin S. H. Rapid amplification of lambda gt11 bacteriophage library inserts from plaques using the polymerase chain reaction (PCR). Biotechniques. 1989 Jun;7(6):568–570. [PubMed] [Google Scholar]
  15. Ezzell R. M., Kenney D. M., Egan S., Stossel T. P., Hartwig J. H. Localization of the domain of actin-binding protein that binds to membrane glycoprotein Ib and actin in human platelets. J Biol Chem. 1988 Sep 15;263(26):13303–13309. [PubMed] [Google Scholar]
  16. Fox J. E. Identification of actin-binding protein as the protein linking the membrane skeleton to glycoproteins on platelet plasma membranes. J Biol Chem. 1985 Oct 5;260(22):11970–11977. [PubMed] [Google Scholar]
  17. Fox J. E., Reynolds C. C., Phillips D. R. Calcium-dependent proteolysis occurs during platelet aggregation. J Biol Chem. 1983 Aug 25;258(16):9973–9981. [PubMed] [Google Scholar]
  18. Ginsburg D., Handin R. I., Bonthron D. T., Donlon T. A., Bruns G. A., Latt S. A., Orkin S. H. Human von Willebrand factor (vWF): isolation of complementary DNA (cDNA) clones and chromosomal localization. Science. 1985 Jun 21;228(4706):1401–1406. doi: 10.1126/science.3874428. [DOI] [PubMed] [Google Scholar]
  19. Hammonds R. G., Jr Protein sequence of DMD gene is related to actin-binding domain of alpha-actinin. Cell. 1987 Oct 9;51(1):1–1. doi: 10.1016/0092-8674(87)90002-x. [DOI] [PubMed] [Google Scholar]
  20. Hartwig J. H., Shevlin P. The architecture of actin filaments and the ultrastructural location of actin-binding protein in the periphery of lung macrophages. J Cell Biol. 1986 Sep;103(3):1007–1020. doi: 10.1083/jcb.103.3.1007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hartwig J. H., Stossel T. P. Isolation and properties of actin, myosin, and a new actinbinding protein in rabbit alveolar macrophages. J Biol Chem. 1975 Jul 25;250(14):5696–5705. [PubMed] [Google Scholar]
  22. Hartwig J. H., Stossel T. P. Structure of macrophage actin-binding protein molecules in solution and interacting with actin filaments. J Mol Biol. 1981 Jan 25;145(3):563–581. doi: 10.1016/0022-2836(81)90545-3. [DOI] [PubMed] [Google Scholar]
  23. Hartwig J. H., Tyler J., Stossel T. P. Actin-binding protein promotes the bipolar and perpendicular branching of actin filaments. J Cell Biol. 1980 Dec;87(3 Pt 1):841–848. doi: 10.1083/jcb.87.3.841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  25. Imamura M., Endo T., Kuroda M., Tanaka T., Masaki T. Substructure and higher structure of chicken smooth muscle alpha-actinin molecule. J Biol Chem. 1988 Jun 5;263(16):7800–7805. [PubMed] [Google Scholar]
  26. Janmey P. A., Hvidt S., Lamb J., Stossel T. P. Resemblance of actin-binding protein/actin gels to covalently crosslinked networks. Nature. 1990 May 3;345(6270):89–92. doi: 10.1038/345089a0. [DOI] [PubMed] [Google Scholar]
  27. Koteliansky V. E., Glukhova M. A., Shirinsky V. P., Smirnov V. N., Bushueva T. L., Filimonov V. V., Venyaminov S. Y. A structural study of filamin, a high-molecular-weight actin-binding protein from chicken gizzard. Eur J Biochem. 1982 Jan;121(3):553–559. doi: 10.1111/j.1432-1033.1982.tb05822.x. [DOI] [PubMed] [Google Scholar]
  28. Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872. doi: 10.1093/nar/12.2.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kwiatkowski D. J., Janmey P. A., Yin H. L. Identification of critical functional and regulatory domains in gelsolin. J Cell Biol. 1989 May;108(5):1717–1726. doi: 10.1083/jcb.108.5.1717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Maizel J. V., Jr, Lenk R. P. Enhanced graphic matrix analysis of nucleic acid and protein sequences. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7665–7669. doi: 10.1073/pnas.78.12.7665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Maruyama K., Ohashi K. Tropomyosin inhibits the interaction of F-actin and filamin. J Biochem. 1978 Oct;84(4):1017–1019. doi: 10.1093/oxfordjournals.jbchem.a132184. [DOI] [PubMed] [Google Scholar]
  32. Messing J., Gronenborn B., Müller-Hill B., Hans Hopschneider P. Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3642–3646. doi: 10.1073/pnas.74.9.3642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Niederman R., Amrein P. C., Hartwig J. Three-dimensional structure of actin filaments and of an actin gel made with actin-binding protein. J Cell Biol. 1983 May;96(5):1400–1413. doi: 10.1083/jcb.96.5.1400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Noegel A. A., Rapp S., Lottspeich F., Schleicher M., Stewart M. The Dictyostelium gelation factor shares a putative actin binding site with alpha-actinins and dystrophin and also has a rod domain containing six 100-residue motifs that appear to have a cross-beta conformation. J Cell Biol. 1989 Aug;109(2):607–618. doi: 10.1083/jcb.109.2.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Nunnally M. H., Powell L. D., Craig S. W. Reconstitution and regulation of actin gel-sol transformation with purified filamin and villin. J Biol Chem. 1981 Mar 10;256(5):2083–2086. [PubMed] [Google Scholar]
  36. Okita J. R., Pidard D., Newman P. J., Montgomery R. R., Kunicki T. J. On the association of glycoprotein Ib and actin-binding protein in human platelets. J Cell Biol. 1985 Jan;100(1):317–321. doi: 10.1083/jcb.100.1.317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Robson B., Suzuki E. Conformational properties of amino acid residues in globular proteins. J Mol Biol. 1976 Nov 5;107(3):327–356. doi: 10.1016/s0022-2836(76)80008-3. [DOI] [PubMed] [Google Scholar]
  38. Rockwell M. A., Fechheimer M., Taylor D. L. A comparison of methods used to characterize gelation of actin in vitro. Cell Motil. 1984;4(3):197–213. doi: 10.1002/cm.970040305. [DOI] [PubMed] [Google Scholar]
  39. Rosenberg S., Stracher A., Lucas R. C. Isolation and characterization of actin and actin-binding protein from human platelets. J Cell Biol. 1981 Oct;91(1):201–211. doi: 10.1083/jcb.91.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Schollmeyer J. V., Rao G. H., White J. G. An actin-binding protein in human platelets. Interactions with alpha-actinin on gelatin of actin and the influence of cytochalasin B. Am J Pathol. 1978 Nov;93(2):433–446. [PMC free article] [PubMed] [Google Scholar]
  42. Seed B. An LFA-3 cDNA encodes a phospholipid-linked membrane protein homologous to its receptor CD2. 1987 Oct 29-Nov 4Nature. 329(6142):840–842. doi: 10.1038/329840a0. [DOI] [PubMed] [Google Scholar]
  43. Shizuta Y., Shizuta H., Gallo M., Davies P., Pastan I. Purification and properties of filamin, and actin binding protein from chicken gizzard. J Biol Chem. 1976 Nov 10;251(21):6562–6567. [PubMed] [Google Scholar]
  44. Stewart M. Introduction to the computer image processing of electron micrographs of two-dimensionally ordered biological structures. J Electron Microsc Tech. 1988 Aug;9(4):301–324. doi: 10.1002/jemt.1060090403. [DOI] [PubMed] [Google Scholar]
  45. Stossel T. P., Chaponnier C., Ezzell R. M., Hartwig J. H., Janmey P. A., Kwiatkowski D. J., Lind S. E., Smith D. B., Southwick F. S., Yin H. L. Nonmuscle actin-binding proteins. Annu Rev Cell Biol. 1985;1:353–402. doi: 10.1146/annurev.cb.01.110185.002033. [DOI] [PubMed] [Google Scholar]
  46. Stossel T. P. From signal to pseudopod. How cells control cytoplasmic actin assembly. J Biol Chem. 1989 Nov 5;264(31):18261–18264. [PubMed] [Google Scholar]
  47. Stossel T. P., Hartwig J. H. Interactions of actin, myosin, and a new actin-binding protein of rabbit pulmonary macrophages. II. Role in cytoplasmic movement and phagocytosis. J Cell Biol. 1976 Mar;68(3):602–619. doi: 10.1083/jcb.68.3.602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Tyler J. M., Anderson J. M., Branton D. Structural comparison of several actin-binding macromolecules. J Cell Biol. 1980 May;85(2):489–495. doi: 10.1083/jcb.85.2.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Wallach D., Davies P. J., Pastan I. Cyclic AMP-dependent phosphorylation of filamin in mammalian smooth muscle. J Biol Chem. 1978 Jul 10;253(13):4739–4745. [PubMed] [Google Scholar]
  51. Wang K., Ash J. F., Singer S. J. Filamin, a new high-molecular-weight protein found in smooth muscle and non-muscle cells. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4483–4486. doi: 10.1073/pnas.72.11.4483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Wang K. Filamin, a new high-molecular-weight protein found in smooth muscle and nonmuscle cells. Purification and properties of chicken gizzard filamin. Biochemistry. 1977 May 3;16(9):1857–1865. doi: 10.1021/bi00628a015. [DOI] [PubMed] [Google Scholar]
  53. Wang K., Singer S. J. Interaction of filamin with f-actin in solution. Proc Natl Acad Sci U S A. 1977 May;74(5):2021–2025. doi: 10.1073/pnas.74.5.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Weihing R. R. Actin-binding and dimerization domains of HeLa cell filamin. Biochemistry. 1988 Mar 22;27(6):1865–1869. doi: 10.1021/bi00406a011. [DOI] [PubMed] [Google Scholar]
  55. Weihing R. R. Purification of a HeLa cell high molecular weight action binding protein and its identification in HeLa cell plasma membrane ghosts and intact HeLa cells. Biochemistry. 1983 Apr 12;22(8):1839–1847. doi: 10.1021/bi00277a015. [DOI] [PubMed] [Google Scholar]
  56. Youssoufian H., McAfee M., Kwiatkowski D. J. Cloning and chromosomal localization of the human cytoskeletal alpha-actinin gene reveals linkage to the beta-spectrin gene. Am J Hum Genet. 1990 Jul;47(1):62–72. [PMC free article] [PubMed] [Google Scholar]
  57. Zhang Z., Lawrence J., Stracher A. Phosphorylation of platelet actin binding protein protects against proteolysis by calcium dependent sulfhydryl protease. Biochem Biophys Res Commun. 1988 Feb 29;151(1):355–360. doi: 10.1016/0006-291x(88)90601-8. [DOI] [PubMed] [Google Scholar]

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