First evidence for the existence of multiple isoforms of bovine serum amyloid-A (apoSAA).
Bovine serum amyloid-A (SAA) was purified from acute-phase high density lipoprotein (HDL) by affinity chromatography and subsequent gel filtration chromatography. The identity of the isolated protein was checked by Western blotting following SDS-urea-PAGE using antisera raised against the purified protein fraction (SAA) and Amyloid A (AA). The antiserum raised against the purified SAA stained Congo red positive regions in the kidney of an AA-amyloidotic cow and reacted on Western blot with an AA-related protein of approximately 14 kDa. Moreover, it immunostained two to three bands, of approximately 14 kDa, present in serum from diseased cows, proportionally to the serum SAA concentration as measured by ELISA. Isoelectric focusing of the purified bovine SAA fraction revealed three major (pI 5.5, 6.0, 6.4) and three minor (pI 4.8, 5.0, 7.3) isoforms and two-dimensional SDS-urea-PAGE confirmed the identity of the major isoforms. Isoelectric focusing of SAA isolated from sera, obtained from cows affected with different diseases, showed a variable ratio of the isoforms. In SAA isolated from serum obtained from a cow suffering from spontaneous AA-amyloidosis only one isoform (pI 4.8) was detectable. It is concluded that the results give first evidence for the existence of multiple isoforms of bovine SAA, occurring in different plasma concentration ratios during different diseases.
Alsemgeest
S P
SP
Department of Pathology, Faculty of Veterinary Medicine, Utrecht University, The Netherlands.
Horadagoda
A
A
Hulskamp-Koch
C K
CK
Tooten
P C
PC
Kim
D H
DH
Niewold
T A
TA
Gruys
E
E
D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D007124:Immunoenzyme Techniques; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D007668:Kidney / Q000737:chemistry; D008297:Male; D000685:Serum Amyloid A Protein / Q000737:chemistry*
D000685:Serum Amyloid A Protein