The Journal of Neuroscience, November 1991, If(11): 3679-3683
Tangles of Alzheimer’s
of Heparan Sulfate Proteoglycan
with the Neurofibrillary
‘Division of Neuropathology,
N. Kalaria,’ Pamela
Institute of Pathology, Case Western
Mountain View, California 94043, and 3Upjohn Company,
Cleveland, Ohio 44106, 2California
The major intracytoplasmic
of paired helical
sible for the formation
sitive sites in NFTs. bFGF binding
NFTs. In the presence
bind to free carboxyl
may play a role in transforming
lesion of Alzheimer’s
(PHFs). The mechanism
of PHFs, as well as their insolubility
heterogeneity, is unknown.
growth factor (bFGF) binds to heparinase-sen-
is due to a heparan
groups in NFT proteins.
We found that ba-
(e.g., Ca*+), HSPG will
soluble proteins into
tangle (NFT’), a distinctive
troversial (Perry, 1987). Two features that make the understand-
ing of the NFT difficult are the complexity
and their insolubility. Although
strated that a soluble form of PHF is solely composed
microtubule-associated protein tau (Greenberg
Lee et al., 1991), NFTs are largely insoluble
antigens distinct from tau, including
et al., 1987; Mulvihill and Perry, 1989) and tropomyosin
loway et al., 1990). The mechanism
poration of these presumably
In a previous study of the role of growth
mation of senile plaques, we found that basic fibroblast
factor (bFGF) binds to NFTs (Perry et al., 1990a,b;
al., 199 1). We now show that bFGF is bound to a heparan
proteoglycan (HSPG) that is also present in NFTs.
that HSPG binds to the free carboxyl
suggest that similar interactions
other proteins to NFTs and render them insoluble.
and mode of formation
(PHFs) in Alzheimer’s
of the neurofibrillary
of their composition
been demon- it has recently
and Davies, 1990;
for the incor-
into NFTs associated is
factors in the for-
groups of the PHFs and
the binding may mediate of
Received Apr. 23, 1991; accepted June 10, 1991.
We thank Sandy Bowen for manuscript preparation and Drs. L. Culp and T.
Rosenheny for advice. This work was supported by NIH Grants AG007552,
AG009287, and AG00795. G.P. is the recipient of NIH Research Career Devel-
opment Award K04-AG00415, and P.C. and M.K. are fellows of the Fogarty
Correspondence should be addressed to George Perry, Ph.D., Division of Neu-
ropathology, Case Western Reserve University, 2085 Adelbert Road, Cleveland,
Copyright 0 199 1 Society for Neuroscience 0270-6474/91/l 13679-05$05.00/O
Materials and Methods
Tissue. The hippocampal
88 yr) and two controls (60 and 64 yr) and placed in methacam (meth-
anol 6 : chloroform 3 : acetic acid l), a fixative that makes no covalent
modification, for 24-48 hr before dehydration and embedding in par-
affin. Tissue from two AD cases (69 and 73 yr) was frozen in liquid
nitrogen-chilled isopentane. We also used a Bouin’s-Hollande
paraffin-embedded sample taken from the frontal cortex of an AD case
(74 yr) taken at biopsy (gift of Dr. S. Chou, Cleveland Clinic Founda-
tion). Paraffin-embedded sections were cut at 6 pm, and frozen sections,
at 10 Wm.
Antibodies. The following antibodies were used: (1) monoclonal
tibody (Ig fraction) to an epitope located in the carboxyl terminal of
bFGF, 48.1 (J. M. Scardina, unpublished
antiserum to an HSPG core protein (Ledbetter et al., 1987; Noonan et
Protein preparations. The following proteins were used: (1) human
bFGF (154 amino acid form) recombinantly
co/i and purified to homogeneity by using anion exchange chromatog-
raphy, (2) heparan sulfate proteoglycan core protein isolated from Engel-
breth-Helm-Swarm tumor (Ledbetter et al., 1985. 1987: Noonan et al.,
1988) and (3) ovalbumin obtained from Sigma.
Preparation of peroxidase-protein
proteins were prepared by coupling horseradish peroxidase (HRP) to
ovalbumin (as a control), bFGF, or HSPG core protein. Briefly, proteins
were dialyzed into 0.9% NaCl in 1 M NaHCO,, pH 9.5, and incubated
for 16 hr at room temperature (RT) with activated peroxidase Pierce
at a ratio adjusted for the molecular weight of the protein to be coupled.
Excess reactive groups were blocked with 0.2 M Tris, and 1% bovine
serum albumin, followed by dialysis in 150 mM NaCl and 50 mM Tris,
Staining. Sections were immunostained
ploying the peroxidase-antiperoxidase
with 3,3’-diaminobenzidine as cosubstrate. Prior to immunostaining,
sections were treated with 3% H,O, in methanol for 30 min to block
endogenous peroxidase. HRP-protein
development for peroxidase activity as above. In some experiments,
tissue sections were treated with 0.2 M NaOH at RT for 1 hr prior to
staining. In control experiments, primary antibodies were omitted and
the results compared with our experimentals. The extent of ligand bind-
ing was evaluated by comparing staining intensities.
Competition, adsorption, and binding experiments. In order to ascer-
tain the specificity of bFGF binding to NFI in sections, we incubated
bFGFat 4°C for 22 hr with either 100 &ml
or 30 &ml protamine and then applied the solutions to the sections
followed by immunostaining with monoclonal antibody to bFGF 48.1.
In other exneriments. we aDDlied 0.5 &ml
various concentrations of bPGF (0.15 ng to 160 &ml
1:4) to the section and incubated for 16 hr at 4°C. In adsorption
periments, the antibody was incubated with antigen for 16 hr at 4°C
before application to tissue sections. Binding of bFGF or HSPG to NFI
without HRP labeling was assessed by incubating the tissue section with
ligand, rinsing with the same buffer used for incubation, hxing with 3.7%
formaldehyde for 5 min, and then immunostaining
to the ligand, but at a greater dilution that does not identify endogenous
gyrus was taken from nine pathologically con-
1985) cases of AD (mean age, 82 yr; range, 73-
observations), and (2) rabbit
produced in Escherichiu
with the antibodies by em-
procedure (Stemberger, 1986)
conjugates were visualized by
heparin, 5 mg/ml polylysine,
bFGF-HRP together with
with an antibody
3660 Perry et al. - HSPG in Neurofibrillary Tangles
Figure 1. bFGF binds to neurofibrillary tangles (arrows), senile plaque
neurites (large arrowheads), and neuropil threads (small arrowheads) as
well as to amyloid deposits (*). In control sections (not shown), bFGF
binding was limited to blood vessels and the few age-related NITS.
Scale bar, 25 Nrn.
Figure 2. NFfs recognized by an antibody to HSPG, note that the
NPTs recognized are intraneuronal. Nucleus is indicated by arrowhead.
Additionally, intraneuronal granules are stained. No staining of NFTs
or granules was seen in the absence of primary antibody. Scale bar, 25
HSPG. In control experiments, the sections were stained with the same
antibody dilution minus the ligand preincubation.
pH and Ca2+ dependence of bFGF and HSPG binding. The influence
of pH on bFGF or HSPG binding to tissue sections was evaluated by
incubating ligand or ligand-HRP at pH 3.1,4.1, 5.0, 6.0, 7.0, 7.6, 8.0,
9.0, 9.9, and 10.8 by using 0.1 M Bis-Tris-HCl from pH 3.1 to 6.0, 0.1
M Tris-HCl for pH 7.0-9.0, and 0.05 M cyclohexylaminopropane
fonic acid-HCl for pH 9.9 and 10.8 as buffers. The dependence
was determined by using CaZ+/EGTA buffers and ratios of 0.3, 0.5,
0.75, 0.9, 1.0, 1.05, 1.25, 1.5, and 3.0 with 2 mM EGTA (Potter and
Gergely, 1975) in 150 mM NaCl and 50 mM Tris-HCl, pH 7.85. Greater
than 1 mM free Ca*+ solutions omitted EGTA.
Protein and sugar mod@ication treatments. Paraffin-embedded
sections were incubated with one of the following: (1) 0.05 U/ml of
either heparinase (0.05 U/ml, ICN, or 1 U/ml, Sigma), heparinase
U/ml, Sigma), heparitinase (0.05 U/ml, ICN or Seikagaku), or a com-
bination of heparinase and heparitinase at RT for 22 hr in 20 mM CaCl,
and 10 mM Tris, pH 7.0 (Schubert et al., 1988). The specificity of
heparinase or heparitinase treatment for heparan removal from sections
was shown by comparison with the effect of 0.0 1 &ml chondroitinase
ABC (ICN) in 20 mM CaCl, and 10 mM Tris, pH 8.0, incubated at RT
for 22 hr or 400 &ml trypsin (Worthington) at 37°C for 10 min in 300
mM NaCl, 20 mM CaCl,, and 50 mM Tris, pH 7.6. As controls for any
residual protease activity in the heparinase preparation, the inhibitor
0.5 mM leupeptin, 1 mM phenylmethylsulfonyl fluoride, or exogenous
protein (1 mg actin/ml) was used. (2) Two percent periodic acid for 16
hr at RT to modify sugar residues (Behrouz et al., 1989); (3) nitrous
acid consisting of 0.24 M sodium nitrite in 1.8 M acetic acid for 90 min
at RT to modify N-linked sulfates (Hirabayashi et al., 1989); (4) 4
U/ml neuraminidase (Sigma. tvoe VI) in 0.2 M acetate. DH 5.4. for 2
hr at 37°C to remove sia!ic &l-residues (Szumanska et al., 1987); (5)
0.1 M ethanolamine with the addition of 20 mM 1 -ethyl-3-(3-dimethyl-
aminopropyl) carbodiimide (carbodiimide) after 10 min at 37°C to
block and convert carboxyl groups to secondary amines (Taniuchi et
al., 1986); (6) 10 mM paraformaldehyde with 60 mM sodium cyano-
borohydride, 10 mM EDTA, and 10 mM phosphate, pH 7.0, for 2 hr at
37°C to block amino groups (Peterson et al., 1979); (7) 400 &ml alkaline
phosphatase (Sigma, type VII-S, bovine intestinal) in 0.1 M Tris, pH
8.0, with 0.0 1 M phenylmethylsulfonyl fluoride for 18 hr followed by 2
U/ml acid phosphatase
pH 5.0, for 3 hr at 37°C (Ueda et al., 1990); or (8) an unembedded
methacam-fixed tissue block of 3 mm thickness treated with 50% hy-
drofluoric acid for 48 hr at 4°C (Mayor et al., 1990) to remove phosphate
groups followed by paraffin embedding.
(Sigma; prostatic, 3200 U/mg) in 0.1 M acetate,
bFGF bound to NFTs (Fig. 1) as well as to amyloid deposits of
senile plaques, an extracellular filamentous lesion, and vessel
walls in both control and AD cases. However, the binding was
more prominent in the AD cases because of the larger number
of NFTs and senile plaques. bFGF binding, regardless as to
whether it was with NFTs, amyloid deposits, or vessels, had the
characteristics shown in Table 1. bFGF showed saturable bind-
ing that appeared to be charge dependent since CaZ+ or other
polycations as well as anions inhibited the binding. Since bFGF
is known to bind to its receptor as well as to HSPG (Rillcin and
Moscatelli, 1989; Yayon et al., 1991), bFGF binding to NFTs
suggested that HSPGs might be present. The blocking of bFGF
binding by treatment of tissue with heparinases or heparitinase
Table 1. Characteristics of bFGF and HSPG binding to NFTs
c9.0 X lfk6 M
bind HSPG (HSPG-HRP).
in neuronal cell bodies containing a nucleus are indicated by arrowheads.
Differential interface contrast microscopy. Scale bar, 25 pm.
Intraneuronal NFIs, as well as extracellular NFl3 (not shown),
NITS and intraneuronal granules recognized
and the finding that an antiserum to an HSPG immunoreacted
with NFTs (Fig. 2) supported bFGF interaction with HSPG in
In order to understand how HSPGs are incorporated into
NFTs, we characterized the HSPG-binding sites in NFTs by
studying the properties of HSPG binding to NFIs (Fig. 3). The
binding characteristics (Tables 1 and 2) suggest that CaZ+ acts
as a bridge between anionic groups on HSPG, for example,
removal of N-linked sulfate groups by nitrous acid or sugar
residue oxidation with periodate blocked HSPG binding to NFTs,
and anions in NFTs. Consistent with the nonspecific nature of
the polycation requirement, protamine (30-100 &ml) can sub-
The Journal of Neuroscience, November 1991, 1 f(11) 3681
stitute for Ca2+. The Ca*+ inhibition of bFGF binding to HSPG
(anion) in NFTs, is probably mediated by the cationic region
of bFGF (Seno et al., 1990). HSPG binding to NFTs is probably
to cationic groups in NFT, candidates include sugar residues of
P-component, a serum glycoprotein (Kalaria and Grahovac,
1990; Kalaria et al., 199 l), HSPG, phosphate, or carboxyl res-
idues in PHF proteins. To distinguish among these possibilities,
we treated sections with a variety of reagents that alter either
sugar residues or amino acid side chains (Table 2). The same
treatments were performed to characterize bFGF-binding sites.
Alteration of sugar residues with reagents such as periodic acid
or nitrous acid, neuraminidase, or heparinases generally in-
creased HSPG binding to NFT. Thus, not only did HSPG not
seem to be bound to sugar residues in NFTs, but the removal
of sugar residues seemed to unmask new binding sites. In con-
trast, sugar-modifying treatments generally reduced bFGF bind-
ing. Moreover, treatment with formaldehyde-cyanoborohy-
dride, to block amino groups, or with phosphatases or
hydrofluoric acid (Mayor et al., 1990), to remove phosphate
groups, did not decrease HSPG binding, indicating that free
amino and phosphate groups in NFTs did not play a role in
HSPG binding. Treatment with carbodiimide, which had no
effect on bFGF binding, blocked HSPG binding, indicating that
free carboxyl groups are required for HSPG binding to NFI.
A perplexing aspect of HSPG binding was that although HSPG
is a protein of the extracellular matrix, HSPG was found in
intraneuronal NFTs. Two alternative possibilities are that the
association of HSPG with NFTs has occurred during postmor-
tem autolysis or after extracellularization of NFTs. Both pos-
sibilities are unlikely because identical findings were obtained
with the tissue obtained at biopsy (not shown), and we found
that neurons with abundant cytoplasm and the nucleus had
HSPG immunoreactivity (Fig. 2) and HSPG-binding sites (Fig.
3). Therefore, HSPG must be associated with intracellular NFTs
It has been well established that extracellular amyloid deposits
of AD associate with molecules present in the extracellular ma-
trix (Snow et al., 1988; Snow and Wight, 1989). However, there
Table 2. Effect of modifying treatments on bFGF and HSPG binding to NFTs
Treatment Modification bFGF
with protease inhibitor
Sialic acid removal
Effect on binding: T, increased; -, no effect; 1, decreased.
3682 Perry et al. * HSPG in Neurofibrillaty Tangles
is considerable evidence that a glycoprotein, possibly a proteo-
glycan, is also associated with NFTs (Mann et al., 1988; Ro-
senblatt et al., 1989; Snow et al., 1989, 1990). The present study
confirms and extends these findings by characterizing the bio-
chemical basis for HSPG interaction with NFT. We have found
that HSPG- and bFGF-binding
these components are likely to be bound to an anion in NFTs
provided by HSPG for bFGF and by carboxyl residues for HSPG.
HSPG incorporation into NFTs occurs intraneuronally
therefore not an artifact resulting from contamination
The presence of HSPG in NFTs may provide an explanation
for two puzzling features of these inclusions:
components and the insolubility.
amyloid filaments in systemic and cerebral amyloidoses,
HSPG deposition is concomitant
mation (Snow et al., 1987a). Although amyloid filaments form
in vitro by self-assembly, in vivo HSPG is likely to be required
as an initiator of amyloid formation because of the low con-
centration of free amyloidogenic subunits (Kisilevsky
1988; Snow and Wight, 1989). HSPG presumably plays this role
because of its high negative charge, which allows it to concen-
trate and protect ligands from proteolysis
uble complexes, as it apparently does for bFGF (Schreiber et
al., 1985; Gospodarowicz and Cheng, 1986; Saksela et al., 1988).
HSPG may play a similar role in the formation
normal filaments of NFTs. It is known
proteins other than bFGF, some of these proteins such as am-
yloid precursor protein (Schubert et al., 1988, 1989; Klier et al.,
1990), the asymmetric form of AChE (Brandon et al., 1985;
Mesulam and Moran, 1987) and P-component (Hamazaki, 1987;
Duong et al., 1989; Kalaria and Grahovac, 1990; Kalaria et al.,
199 1) have been found in NFT’s. Therefore, the compositional
complexity of NFTs may be due to the binding of proteins with
multiple binding sites triggered by the presence of HSPG, a
phenomenon similar to that occurring in the extracellular matrix
in which various proteins self-assemble through multiple hetero-
and homologous bonds (Yurchenco and Johannes, 1990). More-
over, the binding of HSPG to carboxyl groups may explain why
the identified PHF constituents, tau, neurofilaments, and tropo-
myosin, are abundant in glutamate and aspartate residues
(Cleveland et al., 1977; Geisler et al., 1983; Matsumura
198 5). Consistent with this interpretation, antibodies to tau block
HSPG binding (S.L. Siedlak and G. Perry, unpublished obser-
vations). While HSPG has not been detected in enriched PHF
fractions (Sparkman et al., 1990; Lee et al., 199 1; P. Mulvihill
and G. Perry, unpublished observations),
PHF may depend on polycationic bridges not maintained in the
isolation procedure. Although PHF may be primarily a polymer
of tau, PHF formation may require incorporation
components that are rendered insoluble as they are incorporat-
ed. HSPG’s interaction with PHF proteins may provide the key
to the understanding of these mechanisms.
sites are present in NFTs. Both
number of the
HSPG is present in all known
with amyloid filament for-
while making insol-
of the ab-
that HSPG can bind
its association with
Behrouz N, Defossez A, Delacourte A, Hublau P, Mazzucca M (1989)
Alzheimer disease: glycolytic pretreatment dramatically enhances im-
munolabelling of senile plaques and cerebrovascular
stance. Lab Invest 61:576-583.
Brandon E, Maldonado M, Garrido J, Inestrosa NC (1985) Anchorage
of collagen-tailed acetylcholinesterase
mediated by heparan proteoglycans. J Cell Biol 101:985-992.
Cleveland DW, Hawo SY, Kirschner
properties of purified tau factor and the role of tau in microtubule
assembly. J Mol Biol 116:227-247.
Duong T, Pommier EC, Scheibel AD
human serum amyloid P-component
Neuropathol (Berl) 78429437.
Galloway PG, Mulvihill P, Siedlak S, Mijares M, Kawai M, Padgett H,
Kim R, Perry G (1990) Immunochemical
myosin in the neurofibrillary pathology of Alzheimer
Gambetti P, Autilio-Gambetti L, Manetto V, Perry G (1987) Com-
position of paired helical filaments of Alzheimer’s disease as deter-
mined by specific probes. In: Banbury report 27, Molecular
pathology of aging (Davies P, Finch CE, eds), pp 309-320. Cold Spring
Harbor, NY: Cold Spring Harbor Laboratory.
Geisler M, Kaufman E, Fischer S, Plessmann U, Weber K (1983)
Neurofilament architecture combines structural principles of inter-
mediate filaments with carboxyl-terminal extensions
between triplet nroteins. EMBO J 2: 1295-1302.
Gospodarowicz 6, Cheng J (1986) Heparin protects basic and acidic
bFGF from inactivation. J Cell Physiol 126465484.
Greenberg SG. Davies P (1990) A nrenaration
helical ilaments that displays ‘distinct;
gel electrophoresis. Proc Nat1 Acad Sci USA 875827-5831.
Hamazaki H (1987) Ca2+ mediated association of human serum am-
yloid P-component with heparan sulfate and dermatan sulfate. J Biol
Chem 262: 1456-1460.
Hirabayashi Y, Shimizu S, Yamada K (1989) A nitrous acid procedure
as a selective histochemical means of eliminating
glycoconjugates. Histochem J 2 1:687-692.
Kalaria RN, Grahovac I (1990) Serum amyloid P immunoreactivity
in hippocampal tangles, plaques and vessels: implications
across the blood-brain barrier in Alzheimer
Kalaria RN, Galloway PG, Perry G (199 1) Widespread serum amyloid
P immunoreactivity in cortical amyloid deposits and the neurofi-
brillary pathology of Alzheimer’s disease and related disorders. Neu-
ropathol Appl Neurobiol 17: 189-20 1.
Khachaturian ZS (198 5) Diagnosis of Alzheimer’s
rol 42:1097-l 105.
Kisilevsky R, Snow A (1988) The potential significance of sulphated
glycosaminoglycans as a common constituent of all amyloid: or, per-
haps amyloid is not a misnomer. Med Hypotheses 26:23 l-236.
Kher FG, Cole G, Stallcup W, Schubert D (1990) Amyloid p-protein
precursor is associated with extracellular matrix. Brain Res 5 15:336-
Ledbetter SR, Tyrell B, Hassell JR, Horigan EA (1985) Identification
of the precursor protein to basement membrane heparan sulfate pro-
teoglycans. J Biol Chem 260:8 106-8 113.
Ledbetter SR, Fisher LW, Hassell JR (1987) Domain structure of the
basement membrane heparan sulfate proteoglycan. Biochem 26:988-
Lee VMY, Balin BJ, Otvos L, Trojanowski
subunit of paired helical filaments and derivatized forms of normal
Mann DMA, Bonshek RE, Marcyniuk
(1988) Saccharides of senile plaques and neurofibrillary
Alzheimer’s disease. Neurosci Lett 85277-282.
Matsumura F, Yamashiro I, Matsumura
characterization of multiple isoforms of tropomyosin
tured cells. J Biol Chem 260:13851-13859.
Mayor S, Menor AK, Cross GAM, Ferguson MAJ, Dwek RA, Rade-
macher TW (1990) Glycolipid precursors for the membrane anchor
of Trypanosoma brucei variant surface glycoproteins. J Biol Chem
Mesulam MM, Moran MA (1987)
brillary tangles related to age and Alzheimer’s
Mulvihill P, Perry G (1989) Immunoaffinity demonstration that paired
helical filaments of Alzheimer disease share epitopes with neurofi-
laments, MAP2 and tau. Brain Res 484: 150-l 56.
Noonan DM, Horigan EA, Ledbetter SR, Vogeli G, Sasaki M, Yamada
Y, Hassel JR (1988) Identification
ferent domains of the basement membrane heparan sulfate proteo-
glycan. J Biol Chem 263:16379-16387.
MW (1977) Physical chemical
demonstration of tropo-
disease. Am J
increasing in size
of Alzheimer oaired
proteins by polyacrylamide
the N-sulfate of
disease. Brain Res 5 16:
disease. Arch Neu-
JQ (1991) A68: a major
B, Stoddart RW, Torgerson E
S (1985) Purification
from rat cul-
Cholinesterases within neurofi-
disease. Ann Neurol
of cDNA clones encoding dif-
to the extracellular matrix is
The Journal of Neuroscience, November 1991, 7 7(11) 3663 Download full-text
Perry G (ed) (1987)
heimer disease, pp 229. New York Plenum.
Perry G, Siedlak S, Kawai M, Cras P, Mulvihill
Scardina J, Gambetti
threads and senile plaques all contain abundant binding sites for basic
fibroblast growth factor (@FGF). J Neuropathol
Perry G, Siedlak S, Mulvihill P, Kawai M, Cras P, Cordell B, Miriam
Scardina J, Ledbetter S, Greenberg B, Gambetti P (1990b)
rofibrillary tangles contain heparan sulfate proteoglycans. Neurobiol
Peterson DT, Merrick WC, Safer B (1979)
radiolabeled eukaryotic initiation
comnlex formation. J Biol Chem 254:2509-25 16.
Potter JP, Gergely J (1975) The calcium and magnesium binding sites
on troponin and their role in the regulation of myofibrillary adenosine
triphosphate. J Biol Chem 250:4628-4633.
Rifkin DB, Moscatelli D (1989) Recent developments in the cell bi-
ology of basic fibroblast growth factor. J Cell Biol 109: l-6.
Rosenblatt DE, Geula C, Mesulam MM
munostaining in Alzheimer’s disease. Ann Neurol 26:628-634.
Saksela K, Moscatelli D, Sommer A, Ratkin DB (1988)
cell-derived sulfate binds basic fibroblast growth factor and protects
it from nroteolvtic dearadation. J Cell Biol 107:741-75 1.
Schreiber AB, Kenney J; Kowalski WJ, Friesel R, Mehlman T, Maciag
T (1985) Interaction of endothelial cell growth factor with heparin:
characterization by receptor and antibody recognition. Proc Nat1 Acad
Sci USA 82:6138-6142.
Schubert D, Schroeder R, LaCorbiere
Amyloid fl protein is possibly a heparan sulfate proteoglycan core
protein. Science 241:223-224.
Schubert D, LaCorbiere M, Saitoh T, Cole G (1989) Characterization
of an amyloid precursor protein that binds heparin and contains
tyrosine sulfate. Proc Nat1 Acad Sci USA 86:2066-2069.
Seno M, Sasada R, Kurokawa T, Igarashi K (1990) Carboxyl-terminal
structure of basic fibroblast growth factor significantly contributes to
its affinitv for henarin. Eur J Biochem 188:239-245.
Siedlak SL,Cras P, Kawai M, Richey P, Perry G (199 1) Basic fibroblast
growth factor binding is a marker for extracellular
tangles in Alzheimer’s disease. J Histochem Cytochem 39:899-904.
Alterations in the neuronal cytoskeleton in Alz-
P, Cordell B, Miriam
tangles, neuropil P (1990a) Neurofibrillary
Exp Neurol49:3 18.
Binding and release of
factors 2 and 3 during 805 initiation
(1989) Protease nexin I im-
M, Saitoh T, Cole G (1988)
Snow AD, Wight TN
Snow AD, Willmer J, Kisilevsky R (1987a)
cans: a common constituent of all amyloids? Lab Invest 56: 120-l 23.
Snow AD, Willmer J, Kisilevsky R (1987b)
relationship between sulfated proteoglycans and AA amyloid fibrils.
Lab Invest 57:687-698.
Snow AD, Mar H, Nochlin D, Kimata K, Kato M, Suzuki S, Hassell
J, Wight TN (1988) The presence of heparan sulfate proteoglycans
in the neuritic plaques and congophilic
disease. Am J Path01 133:456463.
Snow AD, Lara S, Nochlin D, Wight TN (1989) Cationic dyes reveal
proteoglycans structurally integrated within the characteristic lesions
of Alzheimer’s disease. Acta Neuropathol
Snow AD, Mar H, Nochlin D, Sekiguchi RT, Klimata
Wiaht TN (1990) Earlv accumulation
and in the beta amyloid protein containing
disease and Down’s syndrome. Am J Path01 137: 1253-1270.
Sparkman DR, Hill SJ, White CL (1990)
not the major binding sites for wheat germ and Dofichos fiflorus
agglutinins in the neurofibrillary tangles of Alzheimer’s
Szumanska G, Vorbrodt AW, Mandybur
Lectin histochemistry of plaques and tangles in Alzheimer’s
Acta Neuropathol (Berl) 73: l-l 1.
Taniuchi M, Schweitzer JB, Johnson EM (1986) Nerve growth factor
receptor molecules in rat brain. Proc Nat1 Acad Sci USA 83:1950-
Ueda K, Masliah E, Saitoh T, Bakalis SL, Scoble H, Kosik KS (1990)
Alz-50 recognizes a phosphorylated
Yayon A, Kingsbrun M, Esko JD, Leder P, Ornitz DM
surface, heparin-like molecules are required for binding of basic fi-
broblast growth factor to its high affinity receptor. Cell 64:841-848.
Yurchenco PD, Johannes SC (1990) Molecular
ment membranes. FASEB J 4: 1577-l 590.
(1989) Proteoglycans in the pathogenesis of
disease and other amyloidoses. Neurobiol Aging lo:48 l-
A close ultrastructural
angiopathy in Alzheimer’s
(Berl) 78: 113-l 23.
K, Koike Y,
of henaran sulfate in neurons
lesions in Alzheimer’s
Paired helical filaments are
TI, Wisniewski HM (1987)
epitope of tau protein. J Neurosci
architecture of base-