Biochemical nature and cellular origin of amyloid enhancing factor (AEF) as determined by anti-AEF antibody.

Abstract

Low ionic strength acidic buffer, Sephadex G-200 and Benzamidine-Sepharose (BZ) gel chromatography, have been used for the partial purification of alveolar hydatid cyst (AHC) induced amyloid enhancing factor (AEF). BZ-gel bound AEF (AEF-BZ) demonstrated AEF activity in the mouse bioassay, proteolytic activity against Hide powder azure showed two major and three minor peptides on SDS-PAGE. Pretreatment of AEF-BZ with 10 mM phenylmethylsulphonyl fluoride or 20 mM p-chloromercuribenzoic acid completely abolished its bioactivity in vivo and proteolytic activity in vitro. Polyclonal anti-AEF antibody (AAA) was generated which on passive transfer into mice completely abolished the bioactivity of both casein-induced, or AHC-induced AEF. The AAA absorbed on Sepharose gel conjugated to normal mouse serum developed one common precipitin band between AE and AEF-positive sera from AHC-infected and old retired mice and in immunostaining it bound to the cytoplasmic granular components of a majority of splenic and peritoneal leucocytes from AHC-infected mice. In contrast, only a few normal mouse leucocytes showed positive staining. We suggest that AEF, in all probability, is a serine/thiol protease of leucocyte origin whose intracellular and humoral concentrations increase significantly during amyloidosis. The role of lysosomal proteases and anti-AEF antibody which has been successfully generated for the first time is discussed with reference to the origin of AEF and its presumed biological function in amyloidogenesis.

• Authors: K Alizadeh-Khiavi, Z Ali-Khan
• Journal: British journal of experimental pathology (Br J Exp Pathol) Vol. 69 Issue 5 Pg. 605-19 (Oct 1988) ISSN: 0007-1021 [Print] England
• PMID: 3058197 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Amyloid
• Antibodies
• Chloromercuribenzoates
• Glycoproteins
• amyloid enhancing factor
• Phenylmethylsulfonyl Fluoride
• p-Chloromercuribenzoic Acid
• Peptide Hydrolases

Topics

• Amyloid (analysis)
• Animals
• Antibodies (analysis)
• Chloromercuribenzoates (pharmacology)
• Chromatography, Gel
• Electrophoresis, Polyacrylamide Gel
• Glycoproteins (analysis, antagonists & inhibitors, immunology)
• Immunization, Passive
• Liver (analysis)
• Male
• Mice
• Mice, Inbred BALB C
• Peptide Hydrolases (analysis)
• Phenylmethylsulfonyl Fluoride (pharmacology)
• Spleen (analysis)
• p-Chloromercuribenzoic Acid