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Human MN glycoproteins: Dependence of blood-group and anti-influenza virus activities on their molecular size
Vol. 28, No. 4, 1967
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
DEPENDENCE OF BLOOD-GROUP AND ANTI-INFLUENZA
HUMAN M N GLYCOPROTEINS:
VIRUS ACTIVITIES
ON THEIR
Georg
F.
MOLECULAR SIZE*
Springer
Immunochemistry Department**, Department of Microbiology, Illinois 60201 Evanston,
Received
addition
they
are
potent
possessing
human
red
were
molecules
treatment of
pH 5.5-7.2
of
smaller
weights
very
that
large
molecules
the
have
receptor
M N activity
gentlest of
blood-group been
system;
characterized
preparation has
varying
size
by a series
as
raising any
step
also
of
been
of 30,000
multiples
demonstrable
as the
result
do occur
at
that
manipulation
(7-12). of
slightly
elevated
M N substances.
of
in chemi
related
isolated
above and the and
45°C
frorr
Department
Major more
temperatures,
is
supported
510
or
the
rigorous however
a range
a predominance
molecules differences
molecular tend
to
were
not
minor
chemical
(7,8).
~1-05681 of the American
by the
Harsher
possess
conditions;
a population
outside
gave
large
chemical
from
working
in
blood-group
(7-11).
purification
-k This investigation has been supported by grant Institutes of Health and grant No. 63 G 119 of Immunochemistry
form
M N glycoproteins that
resulted
These
fractionations
isolation
appears
procedures
homogeneous
temperature
during
It
molecules. are
the
isolation
the
physicochemicaily
upon
J;* The
human
and
virus
blood-group
found
disaggregate
alterations
influenza
in
at
which
of
second
receptors
obtained
of such
virus
An
traces
the
(6).
of
substances
(l-5).
recently
preponderance
constitute
influenza
only
ccl Is
We have
of
antigens
as glycoproteins
nature
Hospital and University,
July 5, 1967
The M N erythrocyte
tally
Evanston Northwestern
Susan
the National Heart Association.
Rebecca
Stone
Fund.
a
Vol. 28, No. 4, 1967
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
TABLE 1 --IN VITRO ACTIVITY
OF HUMAN M M AND NN BLOOD-GROUP ANTIGENS:
DEPENDENCE ON MOLECULAR SIZE.*
Antigen
Molecular weight
Smallest amount (pg/ml) agglutination of human cytes by 4 agglutinating
-Human Anti-M
MM Ca 979
12 x
Ca 980
SeraAnti-N
0.05
6 x lo6
3
0.2
5
0.8
NN Ca 825
5.9
x 105
10
Ca 745
1.5
x
50
3.1
x 10 4
lo5
5oo
:k For procedures see (8). ** Homologous, homozygous erythrocytes A*:: Highest blood-group activity out Dr. R.H. Rathan.
We have
now found
and
capacity
the
dependence size
on the
appears
as well
depicted
that to
be one
isolated in
the
Table
--in
inhibit
molecular
as antiviral
on substances are
to
Influenza PR8
1
x lo6
sera
inhibiting 0 erythro-
106
1.8
Ca 1014
completely blood-group doses*;?
of
of
1.
this It
the
important
activities in
vitro
were
can
blood-group
activity
glycoprotein, which
for under
that
511
as measured
by some myxoviruses
isolated
highest
be seen
10
used for blood-group determination. 6 samples kindly furnished by
factors
laboratory
1.5
3,500
hemagglutination
size the
of
virus
the the
virus
i.e.,
determine largest gentle inhibitory
activity. molecules.
conditions capacity
with
shows the
human
a marked
molecular Blood-group The
findings
described increases
above
Vol. 28, No. 4, 1967
with the
molecular most
size
powerful
substantial
and
increase
activity
of
human
red
virus
cell
aqueous
the
these
receptors
(6)
virus
had
~0.5%
of
the
blood-group
and
~1.5%
of
the
smallest.
This
weight
of
which
active
M and
listed
last
last
expressed
in
on a molar
Dependence when
virus
blood-group It
is
molecular
Table
antiviral
determined in
both
Also,
(13).
hemagglutination
to
virus
the
activity
of
by
by a number
of
and
that
the
pH 8 the
the
from
with
highly
of
the
substance
listed
first
activities
with
on molecular
influenza
size
virus
was
B strain
less
for
determination
discussed independent and Mrs.
here
of and
the to
assessment H. Tegtmeyer
Dr.
A.
Dr. of for
8.
data
presented
here
activity
has
submaxillary
viruses
which
became
virus
their expert
Dr.
weights
Rubin,
Wyeth
antiviral
ant
sera.
Bezkorovainy,
molecular
markc
Maryland
that
importance
been
deduced
mucin
following
does
not
a potent
strains
the
it
the
treatment
significantly
inhibitor
when
from
was
of polymerized
ACKNOWLEDGEMENTS to
ant
are
(14,15).
I am grateful
it
Table
haptens
substances if
the
and
in
of
digestion
the
blood-
with
listed
than
striking
in
listed
some damage
of
(Mw 41,000)
influenza
lower
68°C
be
a
comparison
sample
molecules
activity
ovine
influenza
at
to
IO6 fold.
inhibitor
orosomucoid
hemagglutination
more
For
active
indicates
even
Likewise
as measured
by pronase
activity
rabbit
to
is
and
blood-group
with
largest
in
lo4
relation
and
the
was measured
influenza size
is
and
activity
for
of
most
appear
increase
sera.
activity
the
obtained
becomes it
activity
trypsin
inhibit
the basis:
noteworthy
of
The difference
inhibitory
size
decrease with
of
of
cells.
extracted
antiviral
(8,10)
1.
human
been
activity
were
human
weight
by a similar
with
that
latter
macromolecules
in Table
respectively
of
from
had
Its
activities
~5000
N active
which
was 0.5%
molecular
accompanied
as measured
included.
large
isolated
is
material
was
of
yet
size
substances
receptor
influenza
a molecular
glycoproteins
in molecular
phenol
PR8 strain
that
myxovirus
group
with
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
J.
(weight Institute activities.
technical
help.
512
Kirschbaum, average) of
and of
the
Dr.
G. Bernardi
substances
Medical
Research,
I thank
Miss
M.
for Kleintop
of
Vol. 28, No. 4, 1967
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
REFERENCES
I. 2.
Springer, MHkela,
O.,
G.F., and
and Ansell, N.J., Cantell, K., Ann.
Proc. Natl. Med. Exptl.
Acad. Biol.
Sci. U.S. 44, 182 (1958) Fenniae (Helsinki) 36,
366 (1958). 3. 4. 5.
;: 8. 9. 10. 11.
K* , Baranowski, T., Lisowska, E., Morawiecki, A., Romanowska, E., and Strozecka, Arch. Immunol. Terapii Dosw. 1, 15 (1959). Stalder, K., and Springer, G.F., Fed. Proc. 2, 70 (1960). Klenk, E., and Uhlenbruck, G., Z. Physiol. Chem. 319, 151 (1960). Kathan, R.H., Winzler, R.J., and Johnson, C.A., J. Exptl. Med. 113, 37 (1961). Springer, G.F. in Immunchemie, X V Colloquium Germ. Sot. of Physm. Chemists, 1964, pp. 90-112, Springer-Verlag, Berlin, Heidelberg, New York, 1965. Mosbach, Springer, G.F., Nagai, Y., and Tegtmeyer, H., Biochemistry 5, 3254 (1966). Springer, G.F., Fletcher, M., and Pavlovskis, O., X V Colloquium Protides of Biological Fluids, H. Peeters ed., Brugge, 1967, Elsevier, in press. Springer, G.F., Wang, C.S., and Mao, T., in preparation (1967). Bezkorovainy, A., Springer, G.F., and Hotta, K., Biochim. Biophys. Acta l&,
501 (1966). 12.
13. 14.
15.
Morawiecki, A., Biochim. Biophys. Acta 83, 339 (1964). and Biology of Sialic Acids and Related Substances Gottschalk, A., The Chemistry (Camb. Univ. Press, 1960) p. 82. Morawiecki, A., and Lisowska, E., Biochem. and Biophys. Res. Comm. Is, 4, 606 (19651 Whitehead, P.H., Flewett, T.H., Foster, J.R., and Sammons, H.G., Nature 208, 915 (1965).
513
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
DEPENDENCE OF BLOOD-GROUP AND ANTI-INFLUENZA
HUMAN M N GLYCOPROTEINS:
VIRUS ACTIVITIES
ON THEIR
Georg
F.
MOLECULAR SIZE*
Springer
Immunochemistry Department**, Department of Microbiology, Illinois 60201 Evanston,
Received
addition
they
are
potent
possessing
human
red
were
molecules
treatment of
pH 5.5-7.2
of
smaller
weights
very
that
large
molecules
the
have
receptor
M N activity
gentlest of
blood-group been
system;
characterized
preparation has
varying
size
by a series
as
raising any
step
also
of
been
of 30,000
multiples
demonstrable
as the
result
do occur
at
that
manipulation
(7-12). of
slightly
elevated
M N substances.
of
in chemi
related
isolated
above and the and
45°C
frorr
Department
Major more
temperatures,
is
supported
510
or
the
rigorous however
a range
a predominance
molecules differences
molecular tend
to
were
not
minor
chemical
(7,8).
~1-05681 of the American
by the
Harsher
possess
conditions;
a population
outside
gave
large
chemical
from
working
in
blood-group
(7-11).
purification
-k This investigation has been supported by grant Institutes of Health and grant No. 63 G 119 of Immunochemistry
form
M N glycoproteins that
resulted
These
fractionations
isolation
appears
procedures
homogeneous
temperature
during
It
molecules. are
the
isolation
the
physicochemicaily
upon
J;* The
human
and
virus
blood-group
found
disaggregate
alterations
influenza
in
at
which
of
second
receptors
obtained
of such
virus
An
traces
the
(6).
of
substances
(l-5).
recently
preponderance
constitute
influenza
only
ccl Is
We have
of
antigens
as glycoproteins
nature
Hospital and University,
July 5, 1967
The M N erythrocyte
tally
Evanston Northwestern
Susan
the National Heart Association.
Rebecca
Stone
Fund.
a
Vol. 28, No. 4, 1967
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
TABLE 1 --IN VITRO ACTIVITY
OF HUMAN M M AND NN BLOOD-GROUP ANTIGENS:
DEPENDENCE ON MOLECULAR SIZE.*
Antigen
Molecular weight
Smallest amount (pg/ml) agglutination of human cytes by 4 agglutinating
-Human Anti-M
MM Ca 979
12 x
Ca 980
SeraAnti-N
0.05
6 x lo6
3
0.2
5
0.8
NN Ca 825
5.9
x 105
10
Ca 745
1.5
x
50
3.1
x 10 4
lo5
5oo
:k For procedures see (8). ** Homologous, homozygous erythrocytes A*:: Highest blood-group activity out Dr. R.H. Rathan.
We have
now found
and
capacity
the
dependence size
on the
appears
as well
depicted
that to
be one
isolated in
the
Table
--in
inhibit
molecular
as antiviral
on substances are
to
Influenza PR8
1
x lo6
sera
inhibiting 0 erythro-
106
1.8
Ca 1014
completely blood-group doses*;?
of
of
1.
this It
the
important
activities in
vitro
were
can
blood-group
activity
glycoprotein, which
for under
that
511
as measured
by some myxoviruses
isolated
highest
be seen
10
used for blood-group determination. 6 samples kindly furnished by
factors
laboratory
1.5
3,500
hemagglutination
size the
of
virus
the the
virus
i.e.,
determine largest gentle inhibitory
activity. molecules.
conditions capacity
with
shows the
human
a marked
molecular Blood-group The
findings
described increases
above
Vol. 28, No. 4, 1967
with the
molecular most
size
powerful
substantial
and
increase
activity
of
human
red
virus
cell
aqueous
the
these
receptors
(6)
virus
had
~0.5%
of
the
blood-group
and
~1.5%
of
the
smallest.
This
weight
of
which
active
M and
listed
last
last
expressed
in
on a molar
Dependence when
virus
blood-group It
is
molecular
Table
antiviral
determined in
both
Also,
(13).
hemagglutination
to
virus
the
activity
of
by
by a number
of
and
that
the
pH 8 the
the
from
with
highly
of
the
substance
listed
first
activities
with
on molecular
influenza
size
virus
was
B strain
less
for
determination
discussed independent and Mrs.
here
of and
the to
assessment H. Tegtmeyer
Dr.
A.
Dr. of for
8.
data
presented
here
activity
has
submaxillary
viruses
which
became
virus
their expert
Dr.
weights
Rubin,
Wyeth
antiviral
ant
sera.
Bezkorovainy,
molecular
markc
Maryland
that
importance
been
deduced
mucin
following
does
not
a potent
strains
the
it
the
treatment
significantly
inhibitor
when
from
was
of polymerized
ACKNOWLEDGEMENTS to
ant
are
(14,15).
I am grateful
it
Table
haptens
substances if
the
and
in
of
digestion
the
blood-
with
listed
than
striking
in
listed
some damage
of
(Mw 41,000)
influenza
lower
68°C
be
a
comparison
sample
molecules
activity
ovine
influenza
at
to
IO6 fold.
inhibitor
orosomucoid
hemagglutination
more
For
active
indicates
even
Likewise
as measured
by pronase
activity
rabbit
to
is
and
blood-group
with
largest
in
lo4
relation
and
the
was measured
influenza size
is
and
activity
for
of
most
appear
increase
sera.
activity
the
obtained
becomes it
activity
trypsin
inhibit
the basis:
noteworthy
of
The difference
inhibitory
size
decrease with
of
of
cells.
extracted
antiviral
(8,10)
1.
human
been
activity
were
human
weight
by a similar
with
that
latter
macromolecules
in Table
respectively
of
from
had
Its
activities
~5000
N active
which
was 0.5%
molecular
accompanied
as measured
included.
large
isolated
is
material
was
of
yet
size
substances
receptor
influenza
a molecular
glycoproteins
in molecular
phenol
PR8 strain
that
myxovirus
group
with
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
J.
(weight Institute activities.
technical
help.
512
Kirschbaum, average) of
and of
the
Dr.
G. Bernardi
substances
Medical
Research,
I thank
Miss
M.
for Kleintop
of
Vol. 28, No. 4, 1967
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
REFERENCES
I. 2.
Springer, MHkela,
O.,
G.F., and
and Ansell, N.J., Cantell, K., Ann.
Proc. Natl. Med. Exptl.
Acad. Biol.
Sci. U.S. 44, 182 (1958) Fenniae (Helsinki) 36,
366 (1958). 3. 4. 5.
;: 8. 9. 10. 11.
K* , Baranowski, T., Lisowska, E., Morawiecki, A., Romanowska, E., and Strozecka, Arch. Immunol. Terapii Dosw. 1, 15 (1959). Stalder, K., and Springer, G.F., Fed. Proc. 2, 70 (1960). Klenk, E., and Uhlenbruck, G., Z. Physiol. Chem. 319, 151 (1960). Kathan, R.H., Winzler, R.J., and Johnson, C.A., J. Exptl. Med. 113, 37 (1961). Springer, G.F. in Immunchemie, X V Colloquium Germ. Sot. of Physm. Chemists, 1964, pp. 90-112, Springer-Verlag, Berlin, Heidelberg, New York, 1965. Mosbach, Springer, G.F., Nagai, Y., and Tegtmeyer, H., Biochemistry 5, 3254 (1966). Springer, G.F., Fletcher, M., and Pavlovskis, O., X V Colloquium Protides of Biological Fluids, H. Peeters ed., Brugge, 1967, Elsevier, in press. Springer, G.F., Wang, C.S., and Mao, T., in preparation (1967). Bezkorovainy, A., Springer, G.F., and Hotta, K., Biochim. Biophys. Acta l&,
501 (1966). 12.
13. 14.
15.
Morawiecki, A., Biochim. Biophys. Acta 83, 339 (1964). and Biology of Sialic Acids and Related Substances Gottschalk, A., The Chemistry (Camb. Univ. Press, 1960) p. 82. Morawiecki, A., and Lisowska, E., Biochem. and Biophys. Res. Comm. Is, 4, 606 (19651 Whitehead, P.H., Flewett, T.H., Foster, J.R., and Sammons, H.G., Nature 208, 915 (1965).
513