Summary
Crude amyloid enhancing factor (AEF) drastically reduces the pre-amyloid phase on passive transfer and induces amyloid deposition in the recipient mice in 48–120h. We attempted to purify AEF from murine amyloidotic liver and spleen extracts by using gel filtration, preparative sodium dodecyl sulphate-polyacrylamide gel electrophoresis and ion exchange chromatography and isolated a 5.5. kDa peptide. In the mouse bioassay, this peptide induced accelerated splenic AA deposition in a dose-dependent manner. Based on structural, electrophoretic and immunochemical criteria the peptide was identified as ubiquitin. A polyclonal rabbit anti-bovine ubiquitin IgG antibody (RABU) abolished the in vivo AEF activity of crude murine AEF in a dose-dependent manner. Monomeric ubiquitin and its large molecular weight adducts were isolated from crude AEF using cyanogen bromide-activated sepharose conjugated to RABU and size exclusion chromatography methods. These were assayed and were found to possess AEF activity. Furthermore, increased levels of ubiquitin, a phenomenon similar to that of AEF, were detected by immunocytochemistry in mouse peritoneal leucocytes prior to and during amyloid deposition. Since AEF shares a number of biological and functional properties with ubiquitin, we suggest a possible role of ubiquitin as an AEF, and that serum amyloid protein A and ubiquitin, the two reactants generated during inflammatory stress conditions, may converge to induce AA amyloid deposition
Similar content being viewed by others
References
Abankwa GA, Ali-Khan Z (1988) Alveolar hydatid cyst induced amyloid enhancing factor (AEF): physicochemical properties and abolition of AEF activity by serine protease inhibitors. Br J Exp Pathol 69:133–148
Abraham CR, Selkoe DJ, Potter H (1988) Immunochemical identification of the serine protease inhibitor α1-antichymotrypsin in the brain deposits of Alzheimer's disease. Cell 52:487–501
Ali-Khan Z, Quirion R, Robittaile Y, Alizadeh-Khiavi K, Du T (1988) Evidence for increased amyloid enhancing factor activity in Alzheimer brain extract. Acta Neuropathol (Berl) 77:82–90
Alizadeh-Khiavi K, Ali-Khan Z (1988) Biochemical nature and cellular origin of amyloid enhancing factor (AEF) as determined by anti-AEF antibody. Br J Exp Pathol 69:605–619
Alizadeh-Khiavi K, Ali-Khan Z (1990) Possible role of amyloid enhancing factor (ubiquitin) in Alzheimer's disease and reactive amyloidosis. Neurobiol Aging 11:302
Alizadeh-Khiavi K, Normand J, Chronopoulos S, Ali-Khan Z (1991) Alzheimer brain derived ubiqutin has amyloid enhancing factor activity: behavior of ubiquitin during accelerated amyloidogenesis. Acta Neuropathol (Berl) 81:280–286
Axelrad MA, Kisilevsky R (1980) Biological characterizations of amyloid enhancing factor. In: Glenner GG, Pinhoe Costa P, Falcao de Frietas A (eds) Amyloid and amyloidosis. Excerpta Medica, Amsterdam, pp 527–533
Axelrad MA, Kisilevsky R, Willmar J, Cohen SJ, Skinner M (1982) Further characterzation of amyloid-enhancing factor. Lab Invest 47:139–146
Chronopoulos S, Alizadeh-Khiavi K, Normand J, Ali-Khan Z (1991a) Binding of ubiquitin to experimentally induced murine AA amyloid. J Pathol 163:199–203
Chronopoulos S, Lembo P, Alizadeh-Khiavi K, Normand J, Ali-Khan Z (1991 b) Ubiquitin: its potential significance in murine AA amyloidosis. J Pathol (in press)
Cowan DF, Johnson WC (1970) Amyloidosis in the white Peking Duck. I. Relation to social environmental stress. Lab Invest 23:551–555
Du T, Ali-Khan Z (1990) Pathogenesis of secondary amyloidosis in an alveolar hydatid cyst-mouse model: histopathology and immuno/enzyme-histochemical analysis of splenic marginal zone cells during amyloidogenesis. J Exp Pathol 71:313–316
Fried VA, Smith HT, Hildebrandt E, Weiner K (1987) Ubiquitin has intrinsic proteolytic activity: implications for cellular regulation. Proc Natl Sci USA 84:3685–3698
Fuks A, Zucker-Franklin D (1985) Impaired Kupffer cell function precedes development of secondary amyloidosis. J Exp Med 161:1013–1026
Gervais F, Herbert L, Skamena E (1988) Amyloid enhancing factor: production and response in amyloidosis-susceptible and resistant mouse strains. J Leukoc Biol 43:311–316
Glenner GG (1980) Amyloid deposits and amyloidosis. The β-fibrilloses. N Engl J Med 302:1283–1292
Grundke-Iqbal I, Iqbal K, George L, Tung Y, Kim KS, Wisnieski HM (1989) Amyloid protein and neurofibrillary tangles coexist in the same neuron in Alzheimer disease. Proc Natl Acad Sci USA 86:2853–2857
Hall CE, Cross E, Hall O (1960) Amyloidosis and other pathologic changes in mice exposed to chronic stress. Tex Rep Biol Med 18:205–213
Hardt F, Hellung-Larsen P (1972) Transfer amyloidosis: studies on the nature of the amyloid inducing factor in a murine transfer system. Clin Exp Immunol 10:487–492
Hass AL, Bright PM (1985) The immunochemical detection and quantitation of intracellular ubiquitin-protein conjugates. J Biol Chem 260:12464–12473
Hoffman JS, Ericsson LH, Eriksen N, Walsh KA, Benditt EP (1984) Murine tissue amyloid protein AA: NH2-terminal sequence identity with only one of two serum amyloid protein (apoSAA) gene products. J Exp Med 159:641–645
Hol PR, Van Andel ACJ, Van Enderen AM, Draayer J, Gruys E (1985) Amyloid enhancing factor in hamster. Br J Exp Pathol 66:689–697
Ivy GO, Kitani K, Ihara Y (1989) Anomalous accumulation of τ and ubiquitin immunoreactivities in rat brain caused by protease inhibition and by normal aging: a clue to PHF pathogenesis. Brain Res 498:360–365
Kisilevsky R (1983) Amyloidosis: a familial problem in the light of current pathogenetic developments. Lab Invest 49:381–390
Kisilevsky R, Axelrad M, Corbett W, Brunet S, Scott S (1977) The role of inflammatory cells in the pathogenesis of amyloidosis. Lab Invest 37:544–553
Kula RW, Engel WK, Line BR (1977) Scanning for soft-tissue amyloid. Lancet I:92–93
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Matsumoto H, Taniguchi N, Deutsch HF (1984) Isolation characterization, and esterase and CO hydration activities of ubiquitin from bovine erythrocytes. Arch Biochem Biophys 234:426–433
Mayer JR, Arnold J, Laszlo L, Landon M, Lowe J (1991) Ubiquitin in health and disease. Biochem Biophys Acta 1089:141–157
McCubbin WD, Kay CM, Narindrasorasak S, Kisilevsky R (1988) Circular-dichroism studies on two murine serum amyloid A proteins. Biochem J 256:775–783
Meek RL, Eriksen N, Benditt EP (1989) Serum amyloid A in the mouse: sites of uptake and mRNA expression. Am J Pathol 135:411–419
Niewold THA, Hol PR, Van Andel ACJ, Lutz ETG, Gruys E (1987) Enhancement of amyloid induction by amyloid fibril fragments in hamsters. Lab Invest 56:544–549
Niewold THA, Gruys E, Kisilvesky R, Shirahama TS (1991) Fibril amyloid enhancing factor (FACF) — accelerated amyloidosis in the hamster is not dependent on serine esterase activity and mononuclear phagocytosis. Scand J Immunol 34:101–107
Page DL, Glenner GG (1972) Special interaction of wounding in the genesis of ‘spontaneous’ murine amyloidosis. Am J Pathol 67:555–567
Perlmutter DH (1988) Distinct meidators and mechanisms regulate human acute phase gene expression. In: Perdue ML, Feramisco JH, Lindquist S (eds) Stress induced proteins. Liss, New York, pp 257–274
Rechsteiner M (1989) Ubiquitin-mediated pathways for intracellular proteolysis. Annu Rev Cell Biol 3:1–30
Rosenthal CJ, Sullivan L (1979) Serum amyloid A: evidence for its origin in polymorphonuclear leukocytes. J Clin Invest 62:1181–1186
Shirahama T, Miura K, Ju S-T, Kisilevsky R, Gruys E, Cohen AS (1990a) Amyloid enhancing factor-loaded macrophages in amyloid fibril formation. Lab Invest 62:61–68
Shirahama T, Abraham CR, Ju S-T, Miurak K, Cohen AS, Kisilevsky R, Grys E (1990b) Isolation of a 16 KD fraction with extremely high AEF activity. In: Natvig JB et al. (eds) Amyloid and amyloidosis. Kluwer, London, pp 288–291
Silverman SL, Cathcart ES, Skinner M, Cohen AS, Burnett L (1980) A pathogenetic role for polymorphonuclear leukocytes in the synthesis and degradation of SAA protein. In: Glenner GG et al. (eds) Amyloid and amyloidosis. Excerpta Medica, Amsterdam, pp 420–425
Sipe JD, McAdam KPWJ, Uchino F (1978) Biochemical evidence for the biphasic development of experimental amyloidosis. Lab Invest 38:110–114
Skinner M, Stone P, Shirahama T, Connors LH, Calore J, Cohen AS (1986) The association of an elastase with amyloid fibrils. Proc Soc Exp Biol Med 181:211–214
Snow A, Kisilevsky R, Stephens C, Anastassiades T (1987) Characterization of serum and splenic glycosaminoglycans during rapid AA amyloid induction. Qualitative and quantitative analysis. Lab Invest 56:665–675
Stone MJ (1990) Amyloidosis: a final common pathway for protein deposition in tissues. Blood 75:531–545
Turnell W, Sarra R, Glover ID, Baum JO (1986) Secondary structure prediction of human SAA1, Presumptive identification of calcium and lipid binding sites. Mol Biol Med 3:387–407
Varga J, Flinn MSM, Shirahama T, Rogers OG, Cohen AS (1986) The induction of accelerated murine amyloid with human splenic extract. Virchows Arch [B] 51:177–185
Yamamoto KI, Migita S (1985) Complete primary structures of two major murine serum amyloid A proteins deduced from cDNA sequences. Proc Natl Acad Sci USA 82:2915–2919
Yokota T, Ishihara T, Kawano H, Takahashi M, Fujinaga Y, Uchino F (1989) Amyloid enhancing factor (AEF) in the aging mouse. Virchows Arch [A] 414:511–514
Zuckerman SA, Suprenant YM (1986) Simplified micro ELISA for the quantitation of murine serum amyloid A protein. Immunol Methods 92:37–43
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Alizadeh-Khiavi, K., Normand, J., Chronopoulos, S. et al. Amyloid enhancing factor activity is associated with ubiquitin. Vichows Archiv A Pathol Anat 420, 139–148 (1992). https://doi.org/10.1007/BF02358805
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02358805