1 1 . M U C O P O L Y S A C C H A R I D E S IN TISSUE C U L T U R E
365
I. G L Y C O P R O T E I N S
Although studies on the glycoproteins have been reported in increasing numbers during recent years, few investigations have yet been made
in Tissue Culture systems. The analysis of these substances, in which
the polysaccharide and protein components are firmly linked, is
particularly difficult. Separation of the components by preliminary
hydrolysis may not only destroy certain labile sugars, but also introduce
artifacts through the condensation of sugars and amino acids (Montreuil and Scheppler, 1959). Only certain of the above mentioned
techniques are applicable to glycoproteins, or the glycopeptides
derived from them by enzymic digestion, e.g. electrophoresis on
starch (Neuhaus and Letzring, 1958) and chromatography on ion
exchange resins and cellulose (Nolan and Smith, 1962). Both aspartic
acid (Johansen, Marshall and Neuberger, 1960) and glycine (Konno
and Altman, 1958) have been implicated in the linkage of the carbohydrate and protein in glycoproteins from various sources. The
constituent sugars present in hydrolysates of such materials can be
identified by the well known methods, e.g. chromatography and
electrophoresis in borate buffer (Consden and Stanier, 1952). Methods
for the quantitative estimation of sialic acid (see Gottschalk, 1960) and
the application of these to hydrolysates of sialomucoids have been
investigated by Aminoff (1961). Although the sialic acids are readily
destroyed by acid hydrolysis, their isolation from dilute sulphuric acid
hydrolysates by chromatography on ion exchange resins has been
recorded by Svennerholm (1958) and Martensson, Raal and Svennerholm (1958). Methods for the qualitative and quantitative estimation
of these substances by paper chromatography have been described by
Svennerholm and Svennerholm (1958) and by Blix and Lindberg
(1960). Gibbons (1962) has investigated the kinetics of sialic acid
destruction in 0 - 1 N - H 2 S O 4 at 80°C.
The sialic acids also have characteristic infra-red spectra (Gottschalk,
1960).
I I I . T I S S U E C U L T U R E S T U D I E S
A . M U C O P O L Y S A C C H A R I D E D E T E R M I N A T I O N IN
TISSUE C U L T U R E
In addition to the methods summarized in the preceding section,
there are several for the estimation o f total mucopolysaccharides in
solution, e.g. in tissue culture media.
365
I. G L Y C O P R O T E I N S
Although studies on the glycoproteins have been reported in increasing numbers during recent years, few investigations have yet been made
in Tissue Culture systems. The analysis of these substances, in which
the polysaccharide and protein components are firmly linked, is
particularly difficult. Separation of the components by preliminary
hydrolysis may not only destroy certain labile sugars, but also introduce
artifacts through the condensation of sugars and amino acids (Montreuil and Scheppler, 1959). Only certain of the above mentioned
techniques are applicable to glycoproteins, or the glycopeptides
derived from them by enzymic digestion, e.g. electrophoresis on
starch (Neuhaus and Letzring, 1958) and chromatography on ion
exchange resins and cellulose (Nolan and Smith, 1962). Both aspartic
acid (Johansen, Marshall and Neuberger, 1960) and glycine (Konno
and Altman, 1958) have been implicated in the linkage of the carbohydrate and protein in glycoproteins from various sources. The
constituent sugars present in hydrolysates of such materials can be
identified by the well known methods, e.g. chromatography and
electrophoresis in borate buffer (Consden and Stanier, 1952). Methods
for the quantitative estimation of sialic acid (see Gottschalk, 1960) and
the application of these to hydrolysates of sialomucoids have been
investigated by Aminoff (1961). Although the sialic acids are readily
destroyed by acid hydrolysis, their isolation from dilute sulphuric acid
hydrolysates by chromatography on ion exchange resins has been
recorded by Svennerholm (1958) and Martensson, Raal and Svennerholm (1958). Methods for the qualitative and quantitative estimation
of these substances by paper chromatography have been described by
Svennerholm and Svennerholm (1958) and by Blix and Lindberg
(1960). Gibbons (1962) has investigated the kinetics of sialic acid
destruction in 0 - 1 N - H 2 S O 4 at 80°C.
The sialic acids also have characteristic infra-red spectra (Gottschalk,
1960).
I I I . T I S S U E C U L T U R E S T U D I E S
A . M U C O P O L Y S A C C H A R I D E D E T E R M I N A T I O N IN
TISSUE C U L T U R E
In addition to the methods summarized in the preceding section,
there are several for the estimation o f total mucopolysaccharides in
solution, e.g. in tissue culture media.