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. 1981 Nov;78(11):6798–6802. doi: 10.1073/pnas.78.11.6798

Characterization of brevin, a serum protein that shortens actin filaments

David A Harris *,†, James H Schwartz *,†,‡
PMCID: PMC349138  PMID: 6947253

Abstract

We have purified a protein from rabbit serum with a molecular weight of 90,000 that inhibits the polymerization of actin measured viscometrically and that we have named “brevin” (from the Latin breviare, to shorten). From the extent of purification, we estimate that this inhibitor constitutes 0.3% of the total protein in plasma and serum. Brevin is also present in sera from humans and rats. Almost all of the activity in blood is extracellular; only 1% is present in platelets or other cellular elements. Several lines of evidence indicate that brevin is the same protein as the factor described by Fagraeus and Norberg [Fagraeus, A. & Norberg, R. (1978) Curr. Top. Microbiol. Immunol. 82, 1-13] as an actin-depolymerizing factor (ADF). If ADF and brevin are identical, then “ADF” is an inappropriate name because we find that the protein shortens actin filaments without depolymerizing them. Thus, brevin causes little change in the critical concentration of monomeric actin, even though the inhibitor binds to monomeric actin complexed to DNase I-agarose. Binding of brevin to filaments was demonstrated by sedimenting the inhibitor with F-actin. From the amounts of actin and brevin sedimented, and from the lengths of filaments measured by electron microscopy, we calculated that the stoichiometry of binding is one brevin molecule per filament over a wide range of inhibitor concentrations. This stoichiometry suggests that brevin inhibits polymerization by binding at the end of elongating actin filaments, a mechanism similar to that proposed for several intracellular actin-binding proteins and for the cytochalasins. Its abundance suggests that brevin plays an important physiological role in serum, but one not directly concerned with intracellular motility. Therefore its relationship to cytoplasmic actin-binding proteins remains to be determined.

Keywords: actin polymerization, actin-binding protein, plasma protein, cytoplasmic motility

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bamburg J. R., Harris H. E., Weeds A. G. Partial purification and characterization of an actin depolymerizing factor from brain. FEBS Lett. 1980 Nov 17;121(1):178–182. doi: 10.1016/0014-5793(80)81292-0. [DOI] [PubMed] [Google Scholar]
  2. Blikstad I., Markey F., Carlsson L., Persson T., Lindberg U. Selective assay of monomeric and filamentous actin in cell extracts, using inhibition of deoxyribonuclease I. Cell. 1978 Nov;15(3):935–943. doi: 10.1016/0092-8674(78)90277-5. [DOI] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Brenner S. L., Korn E. D. The effects of cytochalasins on actin polymerization and actin ATPase provide insights into the mechanism of polymerization. J Biol Chem. 1980 Feb 10;255(3):841–844. [PubMed] [Google Scholar]
  5. Bretscher A., Weber K. Villin is a major protein of the microvillus cytoskeleton which binds both G and F actin in a calcium-dependent manner. Cell. 1980 Jul;20(3):839–847. doi: 10.1016/0092-8674(80)90330-x. [DOI] [PubMed] [Google Scholar]
  6. Brown S. S., Spudich J. A. Cytochalasin inhibits the rate of elongation of actin filament fragments. J Cell Biol. 1979 Dec;83(3):657–662. doi: 10.1083/jcb.83.3.657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chaponnier C., Borgia R., Rungger-Brändle E., Weil R., Gabbiani G. An actin-destabilizing factor is present in human plasma. Experientia. 1979 Aug 15;35(8):1039–1041. doi: 10.1007/BF01949928. [DOI] [PubMed] [Google Scholar]
  8. Fagraeus A., Norberg R. Anti-actin antibodies. Curr Top Microbiol Immunol. 1978;82:1–13. doi: 10.1007/978-3-642-46388-4_1. [DOI] [PubMed] [Google Scholar]
  9. Goldberg D. J., Harris D. A., Lubit B. W., Schwartz J. H. Analysis of the mechanism of fast axonal transport by intracellular injection of potentially inhibitory macromolecules: evidence for a possible role of actin filaments. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7448–7452. doi: 10.1073/pnas.77.12.7448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gordon D. J., Boyer J. L., Korn E. D. Comparative biochemistry of non-muscle actins. J Biol Chem. 1977 Nov 25;252(22):8300–8309. [PubMed] [Google Scholar]
  11. Griffin E. E., Miller L. L. Effects of hypophysectomy of liver donor on net synthesis of specific plasma proteins by the isolated perfused rat liver. Modulation of synthesis of albumin, fibrinogen, alpha 1-acid glycoprotein, alpha 2-(acute phase)-globulin, and haptoglobin by insulin, cortisol, triiodothyronine, and growth hormone. J Biol Chem. 1974 Aug 25;249(16):5062–5069. [PubMed] [Google Scholar]
  12. Grumet M., Lin S. A platelet inhibitor protein with cytochalasin-like activity against actin polymerization in vitro. Cell. 1980 Sep;21(2):439–444. doi: 10.1016/0092-8674(80)90480-8. [DOI] [PubMed] [Google Scholar]
  13. Harris H. E., Bamburg J. R., Weeds A. G. Actin filament disassembly in blood plasma. FEBS Lett. 1980 Nov 17;121(1):175–177. doi: 10.1016/0014-5793(80)81291-9. [DOI] [PubMed] [Google Scholar]
  14. Harris H. E., Gooch J. An actin depolymerizing protein from pig plasma. FEBS Lett. 1981 Jan 12;123(1):49–53. doi: 10.1016/0014-5793(81)80017-8. [DOI] [PubMed] [Google Scholar]
  15. Hartwig J. H., Stossel T. P. Cytochalasin B and the structure of actin gels. J Mol Biol. 1979 Nov 5;134(3):539–553. doi: 10.1016/0022-2836(79)90366-8. [DOI] [PubMed] [Google Scholar]
  16. Hasegawa T., Takahashi S., Hayashi H., Hatano S. Fragmin: a calcium ion sensitive regulatory factor on the formation of actin filaments. Biochemistry. 1980 Jun 10;19(12):2677–2683. doi: 10.1021/bi00553a021. [DOI] [PubMed] [Google Scholar]
  17. Hitchcock S. E., Carisson L., Lindberg U. Depolymerization of F-actin by deoxyribonuclease I. Cell. 1976 Apr;7(4):531–542. doi: 10.1016/0092-8674(76)90203-8. [DOI] [PubMed] [Google Scholar]
  18. Isenberg G., Aebi U., Pollard T. D. An actin-binding protein from Acanthamoeba regulates actin filament polymerization and interactions. Nature. 1980 Dec 4;288(5790):455–459. doi: 10.1038/288455a0. [DOI] [PubMed] [Google Scholar]
  19. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  20. Lin D. C., Tobin K. D., Grumet M., Lin S. Cytochalasins inhibit nuclei-induced actin polymerization by blocking filament elongation. J Cell Biol. 1980 Feb;84(2):455–460. doi: 10.1083/jcb.84.2.455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. MacLean-Fletcher S., Pollard T. D. Mechanism of action of cytochalasin B on actin. Cell. 1980 Jun;20(2):329–341. doi: 10.1016/0092-8674(80)90619-4. [DOI] [PubMed] [Google Scholar]
  22. Mustard J. F., Perry D. W., Ardlie N. G., Packham M. A. Preparation of suspensions of washed platelets from humans. Br J Haematol. 1972 Feb;22(2):193–204. doi: 10.1111/j.1365-2141.1972.tb08800.x. [DOI] [PubMed] [Google Scholar]
  23. Norberg R., Thorstensson R., Utter G., Fagraeus A. F-Actin-depolymerizing activity of human serum. Eur J Biochem. 1979 Oct 15;100(2):575–583. doi: 10.1111/j.1432-1033.1979.tb04204.x. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Southwick F. S., Stossel T. P. Isolation of an inhibitor of actin polymerization from human polymorphonuclear leukocytes. J Biol Chem. 1981 Mar 25;256(6):3030–3036. [PubMed] [Google Scholar]
  26. Spudich J. A., Watt S. The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971 Aug 10;246(15):4866–4871. [PubMed] [Google Scholar]
  27. Tilney L. G. The role of actin in nonmuscle cell motility. Soc Gen Physiol Ser. 1975;30:339–388. [PubMed] [Google Scholar]
  28. Yin H. L., Zaner K. S., Stossel T. P. Ca2+ control of actin gelation. Interaction of gelsolin with actin filaments and regulation of actin gelation. J Biol Chem. 1980 Oct 10;255(19):9494–9500. [PubMed] [Google Scholar]

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