Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1983 Nov 1;158(5):1600–1614. doi: 10.1084/jem.158.5.1600

Isolation and characterization of senile amyloid--related antigenic substance (SASSAM) from mouse serum. Apo SASSAM is a low molecular weight apoprotein of high density lipoprotein

PMCID: PMC2187135  PMID: 6415208

Abstract

Sera obtained from senescence-accelerated mouse (SAM) and normal mice contained a substance that reacted with antiserum raised against ASSAM, a novel senile amyloid fibril protein isolated from the liver of SAM. This physiological substance, termed "SASSAM" (serum ASSAM-related antigenic substance), migrated to the albumin/prealbumin region in immunoelectrophoresis and the precipitation line formed with anti-ASSAM antiserum was stained positively with both Amide Black 10 B and Oil Red O/Fat Red 7B solutions, thereby suggesting that SASSAM is an alpha lipoprotein. Using Sephadex G-200 gel chromatography, SASSAM was eluted as a high mol wt form of approximately 200,000 daltons. Fractionation of lipoprotein from normal mouse serum by preparative ultra- centrifugation disclosed that SASSAM was found mainly in high density lipoprotein, HDL (the density is between 1.063 and 1.21 g/cm3). The largest amount of SASSAM was found in the HDL2 fraction (the density is between 1.063 and 1.125) and in this fraction SAA was not detected. Furthermore, ASSAM immunoreactivity appeared in the low mol wt proteins (below 10,000 daltons) of apo HDL separated in the buffer containing 8 M urea through Sephadex G-200. In 8 M urea sodium dodecyl sulfate (SDS)- polyacrylamide gel electrophoresis (PAGE), the major components of apolipoproteins in this position, possibly corresponding to apo C proteins, have the same molecular weight, 5,200 daltons, as ASSAM and this component was labeled by anti-ASSAM antiserum after transfer to nitrocellulose paper.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

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

  1. Anders R. F., Natvig J. B., Michaelsen T. E., Husby G. Isolation and characterization of amyloid-related serum protein SAA as a low molecular weight protein. Scand J Immunol. 1975;4(4):397–401. doi: 10.1111/j.1365-3083.1975.tb02642.x. [DOI] [PubMed] [Google Scholar]
  2. Bausserman L. L., Herbert P. N., McAdam K. P. Heterogeneity of human serum amyloid A proteins. J Exp Med. 1980 Sep 1;152(3):641–656. doi: 10.1084/jem.152.3.641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benditt E. P., Eriksen N. Amyloid protein SAA is associated with high density lipoprotein from human serum. Proc Natl Acad Sci U S A. 1977 Sep;74(9):4025–4028. doi: 10.1073/pnas.74.9.4025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown W. V., Baginsky M. L. Inhibition of lipoprotein lipase by an apoprotein of human very low density lipoprotein. Biochem Biophys Res Commun. 1972 Jan 31;46(2):375–382. doi: 10.1016/s0006-291x(72)80149-9. [DOI] [PubMed] [Google Scholar]
  5. Chai C. K. Reticular cell hyperplasia and amyloidosis in a line of mice with low leukocyte counts. Am J Pathol. 1976 Oct;85(1):49–72. [PMC free article] [PubMed] [Google Scholar]
  6. Cornwell G. G., 3rd, Westermark P., Natvig J. B., Murdoch W. Senile cardiac amyloid: evidence that fibrils contain a protein immunologically related to prealbumin. Immunology. 1981 Nov;44(3):447–452. [PMC free article] [PubMed] [Google Scholar]
  7. Costa P. P., Figueira A. S., Bravo F. R. Amyloid fibril protein related to prealbumin in familial amyloidotic polyneuropathy. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4499–4503. doi: 10.1073/pnas.75.9.4499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  9. Eriksen N., Benditt E. P. Isolation and characterization of the amyloid-related apoprotein (SAA) from human high density lipoprotein. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6860–6864. doi: 10.1073/pnas.77.11.6860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Glenner G. G. Amyloid deposits and amyloidosis. The beta-fibrilloses (first of two parts). N Engl J Med. 1980 Jun 5;302(23):1283–1292. doi: 10.1056/NEJM198006053022305. [DOI] [PubMed] [Google Scholar]
  11. Glenner G. G., Ein D., Eanes E. D., Bladen H. A., Terry W., Page D. L. Creation of "amyloid" fibrils from Bence Jones proteins in vitro. Science. 1971 Nov 12;174(4010):712–714. doi: 10.1126/science.174.4010.712. [DOI] [PubMed] [Google Scholar]
  12. Gorevic P. D., Levo Y., Frangione B., Franklin E. C. Polymorphism of tissue and serum amyloid A (AA and SAA) proteins in the mouse. J Immunol. 1978 Jul;121(1):138–140. [PubMed] [Google Scholar]
  13. Herbert P. N., Windmueller H. G., Bersot T. P., Shulman R. S. Characterization of the rat apolipoproteins. I. The low molecular weight proteins of rat plasma high density lipoproteins. J Biol Chem. 1974 Sep 25;249(18):5718–5724. [PubMed] [Google Scholar]
  14. Higuchi K., Matsumura A., Honma A., Takeshita S., Hashimoto K., Hosokawa M., Yasuhira K., Takeda T. Systemic senile amyloid in senescence-accelerated mice. A unique fibril protein demonstrated in tissues from various organs by the unlabeled immunoperoxidase method. Lab Invest. 1983 Feb;48(2):231–240. [PubMed] [Google Scholar]
  15. Hoffman J. S., Benditt E. P. Changes in high density lipoprotein content following endotoxin administration in the mouse. Formation of serum amyloid protein-rich subfractions. J Biol Chem. 1982 Sep 10;257(17):10510–10517. [PubMed] [Google Scholar]
  16. Isersky C., Ein D., Page D. L., Harada M., Glenner G. G. Immunochemical cross-reactions of human amyloid proteins with human immunoglobulin light polypeptide chains. J Immunol. 1972 Feb;108(2):486–493. [PubMed] [Google Scholar]
  17. Jackson R. L., Baker H. N., Gilliam E. B., Gotto A. M., Jr Primary structure of very low density apolipoprotein C-II of human plasma. Proc Natl Acad Sci U S A. 1977 May;74(5):1942–1945. doi: 10.1073/pnas.74.5.1942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. LaRosa J. C., Levy R. I., Herbert P., Lux S. E., Fredrickson D. S. A specific apoprotein activator for lipoprotein lipase. Biochem Biophys Res Commun. 1970 Oct 9;41(1):57–62. doi: 10.1016/0006-291x(70)90468-7. [DOI] [PubMed] [Google Scholar]
  19. Lavie G., Zucker-Franklin D., Franklin E. C. Degradation of serum amyloid A protein by surface-associated enzymes of human blood monocytes. J Exp Med. 1978 Oct 1;148(4):1020–1031. doi: 10.1084/jem.148.4.1020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lavie G., Zucker-Franklin D., Franklin E. C. Elastase-type proteases on the surface of human blood monocytes: possible role in amyloid formation. J Immunol. 1980 Jul;125(1):175–180. [PubMed] [Google Scholar]
  21. Levin M., Pras M., Franklin E. C. Immunologic studies of the major nonimmunoglobulin protein of amyloid. I. Identification and partial characterization of a related serum component. J Exp Med. 1973 Aug 1;138(2):373–380. doi: 10.1084/jem.138.2.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Linke R. P., Sipe J. D., Pollock P. S., Ignaczak T. F., Glenner G. G. Isolation of a low-molecular-weight serum component antigenically related to an amyloid fibril protein of unknown origin. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1473–1476. doi: 10.1073/pnas.72.4.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Marhaug G., Husby G. Characterization of human amyloid-related protein SAA as a polymorphic protein: association with albumin and prealbumin in serum. Clin Exp Immunol. 1981 Jul;45(1):97–106. [PMC free article] [PubMed] [Google Scholar]
  24. McAdam K. P., Sipe J. D. Murine model for human secondary amyloidosis: genetic variability of the acute-phase serum protein SAA response to endotoxins and casein. J Exp Med. 1976 Oct 1;144(4):1121–1127. doi: 10.1084/jem.144.4.1121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Scanu A. M., Edelstein C. Solubility in aqueous solutions of ethanol of the small molecular weight peptides of the serum very low density and high density lipoproteins: relevance to the recovery problem during delipidation of serum lipoproteins. Anal Biochem. 1971 Dec;44(2):576–588. doi: 10.1016/0003-2697(71)90247-8. [DOI] [PubMed] [Google Scholar]
  26. Scanu A. Forms of human serum high density lipoprotein protein. J Lipid Res. 1966 Mar;7(2):295–306. [PubMed] [Google Scholar]
  27. Scheinberg M. A., Cathcart E. S., Eastcott J. W., Skinner M., Benson M., Shirahama T. The SJL/J mouse: a new model for spontaneous age-associated amyloidosis. I. Morphologic and immunochemical aspects. Lab Invest. 1976 Jul;35(1):47–54. [PubMed] [Google Scholar]
  28. Shirahama T., Skinner M., Westermark P., Rubinow A., Cohen A. S., Brun A., Kemper T. L. Senile cerebral amyloid. Prealbumin as a common constituent in the neuritic plaque, in the neurofibrillary tangle, and in the microangiopathic lesion. Am J Pathol. 1982 Apr;107(1):41–50. [PMC free article] [PubMed] [Google Scholar]
  29. Skinner M., Cohen A. S. The prealbumin nature of the amyloid protein in familial amyloid polyneuropathy (FAP)-swedish variety. Biochem Biophys Res Commun. 1981 Apr 30;99(4):1326–1332. doi: 10.1016/0006-291x(81)90764-6. [DOI] [PubMed] [Google Scholar]
  30. Skogen B., Børresen A. L., Natvig J. B., Berg K., Michaelsen T. E. High-density lipoprotein as carrier for amyloid-related protein SAA in rabbit serum. Scand J Immunol. 1979;10(1):39–45. doi: 10.1111/j.1365-3083.1979.tb01332.x. [DOI] [PubMed] [Google Scholar]
  31. Skogen B., Natvig J. B., Bøorresen A. L., Berg K. Degradation of amyloid-related serum protein SAA by a component present in rabbit and human serum. Scand J Immunol. 1980;11(6):643–648. doi: 10.1111/j.1365-3083.1980.tb00033.x. [DOI] [PubMed] [Google Scholar]
  32. Skogen B., Natvig J. B. Degradation of amyloid proteins by different serine proteases. Scand J Immunol. 1981 Oct;14(4):389–396. doi: 10.1111/j.1365-3083.1981.tb00579.x. [DOI] [PubMed] [Google Scholar]
  33. Skogen B., Thorsteinsson L., Natvig J. B. Degradation of protein SAA to an AA-like fragment by enzymes of monocytic origin. Scand J Immunol. 1980;11(5):533–540. doi: 10.1111/j.1365-3083.1980.tb00021.x. [DOI] [PubMed] [Google Scholar]
  34. Sletten K., Westermark P., Natvig J. B. Senile cardiac amyloid is related to prealbumin. Scand J Immunol. 1980;12(6):503–506. doi: 10.1111/j.1365-3083.1980.tb00098.x. [DOI] [PubMed] [Google Scholar]
  35. Swaney J. B., Reese H., Eder H. A. Polypeptide composition of rat high density lipoprotein: characterization by SDS-gel electrophoresis. Biochem Biophys Res Commun. 1974 Jul 24;59(2):513–519. doi: 10.1016/s0006-291x(74)80010-0. [DOI] [PubMed] [Google Scholar]
  36. Swank R. T., Munkres K. D. Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate. Anal Biochem. 1971 Feb;39(2):462–477. doi: 10.1016/0003-2697(71)90436-2. [DOI] [PubMed] [Google Scholar]
  37. THUNG P. J. Senile amyloidosis in mice. Gerontologia. 1957;1(5):259–279. doi: 10.1159/000210704. [DOI] [PubMed] [Google Scholar]
  38. Takeda T., Hosokawa M., Takeshita S., Irino M., Higuchi K., Matsushita T., Tomita Y., Yasuhira K., Hamamoto H., Shimizu K. A new murine model of accelerated senescence. Mech Ageing Dev. 1981 Oct;17(2):183–194. doi: 10.1016/0047-6374(81)90084-1. [DOI] [PubMed] [Google Scholar]
  39. Takeshita S., Hosokawa M., Irino M., Higuchi K., Shimizu K., Yasuhira K., Takeda T. Spontaneous age-associated amyloidosis in senescence-accelerated mouse (SAM). Mech Ageing Dev. 1982 Sep;20(1):13–23. doi: 10.1016/0047-6374(82)90070-7. [DOI] [PubMed] [Google Scholar]
  40. Tawara S., Araki S., Toshimori K., Nakagawa H., Ohtaki S. Amyloid fibril protein in type I familial amyloidotic polyneuropathy in Japanese. J Lab Clin Med. 1981 Dec;98(6):811–822. [PubMed] [Google Scholar]
  41. 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]
  42. Westermark P. The heterogeneity of protein AA in secondary (reactive)systemic amyloidosis. Biochim Biophys Acta. 1982 Feb 4;701(1):19–23. doi: 10.1016/0167-4838(82)90306-5. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES