Age-dependent degradation of amyloid precursor protein in the post-mortem mouse brain cortex.
We have examined the degradation of amyloid precursor protein (APP) in the brain cortex of adult (24 +/- 2) and old (58 +/- 2) mice at different post-mortem time intervals (0, 1.5, 3, 6, 12 and 24 h). The brain cortex extract was prepared and processed for immunoblotting using antibodies against N-terminal 47-62 amino acids (Asp29) and central 301-316 amino acids containing Kunitz protease inhibitor (KPI) domain (Asp45) of APP. Asp29 (N-terminal) recognizes two bands of 140 and 112 kDa. The amount of 140 kDa is relatively higher in adult than old. The level of 112 kDa is 1.6 times lower in adult than old. It shows no remarkable change with varying post-mortem time. On the other hand, Asp45 (KPI) detects two bands of 110 and 116 kDa. While 116 kDa disappears rapidly after death of the animal, 110 kDa shows no remarkable change with different post-mortem periods. Further incubation of the disrupted tissue at 4 degrees C for 24 h and immunoblot analysis with Asp29 (N-terminal) shows 112 kDa in both ages but 58.5 kDa in adult and 70 kDa in old only. Analysis with Asp45 (KPI) shows only 54 kDa which increases after 3 h in adult but decreases significantly after 1.5 h and becomes undetectable at 24 h in old. Thus the present findings indicate that APP is degraded in a precise pattern and it depends on cellular intactness, post-mortem period and age of the animal.
Asaithambi
A
A
Biochemistry & Molecular Biology Laboratory, Centre of Advanced Study in Zoology, Banaras Hindu University, Varanasi, India.
Mukherjee
S
S
Thakur
M K
MK
D000367:Age Factors; D016564:Amyloid beta-Protein Precursor / Q000378:metabolism*; D000818:Animals; D002457:Cell Extracts / Q000737:chemistry / Q000276:immunology; D002540:Cerebral Cortex / Q000737:chemistry* / Q000473:pathology; D015151:Immunoblotting; D008247:Lysosomes / Q000201:enzymology; D008297:Male; D051379:Mice; D008806:Mice, Inbred AKR; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000378:metabolism; D020033:Protein Isoforms; D013997:Time Factors
D016564:Amyloid beta-Protein Precursor; D002457:Cell Extracts; D020033:Protein Isoforms; D010447:Peptide Hydrolases