Identification of the Mucopolysaccharides in Catecholamine‐Containing Subcellular Particle Fractions from Various Rat, Cat and Ox Tissues

Abstract

Subcellular fractions containing adrenergic vesicles were obtained from bovine adrenal medulla and splenic nerve, rat and cat heart, and rat spleen. The fractions were washed free from lipids, digested with papain and the mucopolysaccharides (MPSs) then precipitated from the supernatant with ethanolic potassium acetate. The mpsswere identified by several different methods--microelectrophoresis on cellulose acetate in various media, cellulose column chromatography using elution solvents of increasing ionic strength, and treatment with specific enzymes followed by electrophoresis or column chromatography. The MPS precipitate from all the sources investigated contained dermatan sulphate or a dermatan sulphate-chondroitin sulphate hybrid. in addition, the precipitate from rat spleen was found to contain chondroitin sulphate. Heparan sulphate was found in the precipitates from rat heart and spleen and hyaluronic acid in that from bovine splenic nerve. The finding of sulphomucopolysaccharides in the adrenergic vesicles, probably in a complex with protein, raises the question of the functional significance of such complexes. They might, by analogy with the ion-exchange function of the heparin-protein complex in mast cell granules, play a role in the storage and release of the amines.

The effect of surgical sympathectomy and of neonatal treatment with 6‐hydroxydopamine and guanethidine on particle‐bound noradrenaline and 35S‐sulphomucopolysaccharides

“…Accordingly, the parallel gradient distribution of 3H-NA and 35S in the present experiments is in agreement with our previous proposals that noradrenergic vesicles contain SMPSs (Aborg et al 1972, Blaschke et al 1976). The concomitant reduction of the accumulation of 3H-NA and 35S in the noradrenergic vesicle fractions after sympathectomy further supports this idea.…”

“…The density gradient distribution of the "-NA containing subcellular particles from homogenized innervated and denervated atria was identical, in accordance with the assumption that in both cases the 3H-NA is localized to noradrenergic vesicles. About 70% of the total 35S in noradrenergic vesicle fractions from cat atria has been found to be present in non-lipid compounds (Blaschke et al 1976).…”

“…The non-lipid "S-labelled compounds in the P,,, ,,oo fractions correspond presumably to SMPSs and possibly sulphated glycoproteins. 35S-sulphate has been shown to be incorporated into these macromolecules in biogenic-amine-storing organelles from the adrenals (Fillion et al 1971, Geissler et al 1977, from sympathetic nerves (Aborg et al 1972, Blaschke et al 1976) and from the brain (Robinson & Green 1962, Pycock et al 1975. In both the 6-OH-DA and the guanethidine-treated rats, the rate of 35S-incorporation into SMPSs of the PIz6 w,, fractions was in most organs considerably less depressed than the 3H-NA uptake-in contrast to the parallel changes of the two processes in atrial tissue after stellate ganglionectomy.…”

“…The parallel distribution curves of particle-bound amines and Y 3 on density gradients indicated that SMPSs might occur in the amine-storing vesicles. The SMPSs were identified chemically as dermatan-, chondroitin-and heparan sulphates or hybrid forms thereof, the SMPS pattern varying somewhat between the tissues studied (Blaschke et al 1976).…”

Biogenesis of Chromaffin Granules: Incorporation of Sulfate into Chromogranin B and into a Proteoglycan

“…In addition, several proteins unrelated to this component are present: dopamine p-hydroxylase (Hortnagl et al, 1974), glycoprotein I11 . enkephalin precursors (Viveros et al, 1979;Udenfriend and Kilpartrick, 1983;Fischer-Colbrie and Frischenschlager, 1985), proteoglycans (Margolis and Margolis, 1973;Blaschke et al, 1976;Geissler et al, 1977;Kiang et al, 1982), and chromogranins B Fischer-Colbrie and Frischenschlager, 1985). These latter proteins, which were characterized recently, represent a family of immunologically related components focusing at a p l of 5.2.…”