Biochemical properties of endogenous presenilin 1 and presenilin 2 in cultured human B-lymphocytes.
BACKGROUND
Presenilin 1 (PS1) and presenilin 2 (PS2) are membranous proteins involved in the pathology of Alzheimer's disease. The development of specific therapies targeted at PS1 or PS2 requires the determination of biochemical properties of presenilins. Hence, in this study we analyzed the hydrophobic and ionic properties of endogenous presenilins.


METHODS
Lysates of immortalized human B-lymphocytes were used as a source of endogenous presenilins. The presence of 3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propanesulfonate (CHAPSO) detergent in lysates favored preservation of PS1 and PS2 native protein complexes. We compared Kyte-Doolittle hydropathicity profiles and hydrophobic interactions of PS1 and PS2 with phenyl-agarose. We also compared the ionic properties of presenilins using anion-exchange chromatography.


RESULTS
The hydropathicity profiles of PS1 and PS2 revealed similarly located hydrophobic regions and more hydrophobic region in the C-terminal fragment of PS2. However, both PS1 and PS2 under physiological conditions showed no interactions with phenyl-agarose. Despite similar predicted isoelectric points, PS1 and PS2 exhibited different ionic behavior during anion-exchange chromatography.


CONCLUSIONS
The different than expected hydrophobic and ionic behavior of PS1 and PS2 may be caused by interactions with other proteins present in complexes formed by endogenous presenilins. The observed difference in ionic properties of PS1 and PS2 can be further explained assuming that PS1 and PS2 form complexes with different sets of proteins. The composition of such variegated PS1 and PS2 complexes can be explored using a proteomic approach. The difference in PS1 and PS2 ionic behavior can be used for purification of endogenous PS1 from PS2, which has not yet been achieved by any other means.
Bojarski
Lukasz
L
Laboratory of Neurodegeneration, International Institute of Molecular and Cell Biology, Warsaw, Poland.
Lewandowicz
Andrzej
A
Blazejczyk
Magdalena
M
Sobczak
Adam
A
Kuznicki
Jacek
J
Wojda
Urszula
U
D000544:Alzheimer Disease / Q000378:metabolism / Q000473:pathology*; D001402:B-Lymphocytes / Q000378:metabolism*; D002478:Cells, Cultured; D002793:Cholic Acids / Q000737:chemistry / Q000494:pharmacology; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007477:Ions; D053764:Presenilin-1 / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D053766:Presenilin-2 / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*
D002793:Cholic Acids; D007477:Ions; D053764:Presenilin-1; D053766:Presenilin-2; C048531:chapso