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.

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 10B and Oil Red O/Fat Red 7B solutions, suggesting that SASSAM is an .alpha. lipoprotein. Using Sephadex G-200 gel chromatography, SASSAM was eluted as a high-MW form of .apprx. 200,000 daltons. Fractionation of lipoprotein from normal mouse serum by preparative ultracentrifugation disclosed that SASAM 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. ASSAM immunoreactivity appeared in the low-MW proteins (< 10,000) of apo-HDL separated in the buffer containing 8 M urea through Sephadex G-200. In 8 M urea sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the major components of apolipoproteins in this position, possibly corresponding to apo C proteins, have the same MW, 5200 as ASSAM, and this component was labeled by anti-ASSAM antiserum after transfer to nitrocellulose paper.