L. Delafournierebessueille et al., PURIFICATION AND SPECTROSCOPIC CHARACTERIZATION OF BETA-AMYLOID PRECURSOR PROTEIN FROM PORCINE BRAINS, European journal of biochemistry, 250(3), 1997, pp. 705-711
Soluble and membrane-bound isoforms of beta-amyloid protein precursor
(APP) of Alzheimer's disease were extracted and purified from porcine
brains. At least three types of soluble APP and membrane-bound APP wit
h different molecular masses, ranging from 86 kDa to 116 kDa, were obt
ained. CD and infrared spectroscopies were used to determine the overa
ll secondary-structure content of APP. The infrared spectra of soluble
and membrane-bound APP (in dry and hydrated states) were similar in t
he amide-I and amide-II regions, suggesting that the overall secondary
structures of the soluble and membrane isoforms were roughly identica
l. The amide-I band is composed of at least five component bands, loca
ted at 1694, 1674, 1652, 1637 and 1618 cm(-1) for soluble APP, and loc
ated at 1687, 1674: 1652, 1637 and 1614-1606 cm(-1) for membrane-bound
APP, as evidenced by their respective second-derivative infrared spec
tra. The 1651-1652-cm(-1) band was associated with alpha-helix structu
res, while two types of beta-sheet structures are evidenced by two cha
racteristic pairs of component bands. The 1674-cm(-1) and 1637-cm(-1)
bands for soluble APP and membrane-bound APP were tentatively associat
ed to beta-sheet structures. The second pair of bands, located at 1694
cm(-1) and at 1618 cm(-1) for soluble APP and at 1687 cm(-1) and 1614
-1606 cm(-1) for membrane-bound APP, were associated with intermolecul
ar beta-sheet structures or aggregated strands, as confirmed by heat d
enaturation. CD spectra indicated the presence of alpha-helix structur
es in soluble and membrane-bound APP. The secondary-structure content,
estimated from CD spectra, was about 40-45% alpha-helix and 15-20% be
ta-sheet structures for soluble and membrane-bound APP.