Detection of distinct isoform patterns of the beta-amyloid precursor protein in human platelets and lymphocytes.
Cerebral deposition of the amyloid beta-protein (A beta P), approximately 40 residue fragment of the integral membrane protein, beta-amyloid precursor protein (beta APP), has been implicated as the probable cause of some cases of familial Alzheimer's disease (AD). The parallels between A beta P deposition in AD and the deposition of certain plasma proteins in systemic amyloid diseases has heightened interest in the analysis of beta APP in circulating cells and plasma. Here, we describe distinct isoform patterns of beta APP in peripheral platelets and lymphocytes. PCR-mediated amplification of mRNA from purified platelets demonstrated the expression of all three major beta APP transcripts (beta APP770,751,695). The full-length, approximately 140 kDa form of beta APP751,770 was detected in membranes of resting and activated platelets but very little immature, approximately 122 kDa beta APP751,770 was found, suggesting a different processing of beta APP in platelets than that described in a variety of cultured cells and tissues. Platelets stimulated with thrombin, calcium ionophore, or collagen released the soluble, carboxyl-truncated form of beta APP (protease nexin-II), but no evidence for the shedding of full-length beta APP associated with platelet microparticles was found, in contrast to previous reports. As a positive control marker for microparticles, the fibrinogen receptor subunit, GPIIIa, was readily detected in platelet releasates. Resting and activated platelets contained similar amounts of the approximately 10 kDa carboxyl terminal beta APP fragment that is retained in platelet membranes following the constitutive cleavage of protease nexin-II. Nonstimulated peripheral B and T lymphocytes contained small amounts of membrane-associated mature and immature beta APP751,770. The potentially amyloidogenic full-length beta APP molecules present in circulating platelets and lymphocytes but not in microparticles could serve as a source of the microvascular A beta P deposited during aging and particularly in AD.
Schlossmacher
M G
MG
Center for Neurologic Diseases, Harvard Medical School, Brigham and Women's Hospital, Boston, MA 02115.
Ostaszewski
B L
BL
Hecker
L I
LI
Celi
A
A
Haass
C
C
Chin
D
D
Lieberburg
I
I
Furie
B C
BC
Furie
B
B
Selkoe
D J
DJ
D000328:Adult; D000595:Amino Acid Sequence; D016564:Amyloid beta-Protein Precursor / Q000097:blood*; D001772:Blood Cell Count; D001792:Blood Platelets / Q000737:chemistry*; D015153:Blotting, Western; D006801:Humans; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000097:blood; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis
D016564:Amyloid beta-Protein Precursor; D010446:Peptide Fragments; D012333:RNA, Messenger