Composition and biogenesis of complex carbohydrates of ox adrenal chromaffin granules

Geissler, Dietmar; Martinek, A.; Margolis, Renée K.; Skrivanek, J. A.; Ledeen, Robert W.; König, Paul; Winkler, Hans

doi:10.1016/0306-4522(77)90023-9

Cited by 64 publications

(33 citation statements)

References 40 publications

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“…A feature that distinguishes such vesicles from those more proximal in the secretory pathway is their higher contents of glycosphingolipids (e.g. the ganglioside GM3 in secretory granules [33]), whose synthesis is completed in the TGN and which are confined to the lumenal membrane leaflet [34]. Glycosphingolipids are characterized by bulky polar headgroups, saturated fatty acyl chains and the trans ‐unsaturated sphingosine, all of which contribute to an inverted‐cone shape that is better compatible with positive than negative membrane curvature.…”

Section: Discussionmentioning

confidence: 99%

Cholesterol is Required for the Formation of Regulated and Constitutive Secretory Vesicles from the trans‐Golgi Network

Wang¹,

Thiele

Huttner

2000

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We studied the role of cholesterol in regulated protein secretion in neuroendocrine cells by manipulating the cholesterol content of AtT-20 cells. Depletion of cellular cholesterol levels caused a reversible block of immature secretory granule biogenesis at the level of the trans-Golgi-network, whereas increased cholesterol levels promoted immature secretory granule formation. Cholesterol depletion also blocked the formation of constitutive secretory vesicles, but did not inhibit the transport between the endoplasmic reticulum and the Golgi complex. Our results indicate that the assembly of cholesterol-based lipid microdomains is required for the biogenesis of both regulated and constitutive secretory vesicles from the trans-Golgi-network in neuroendocrine cells.

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Section: Discussionmentioning

confidence: 99%

Cholesterol is Required for the Formation of Regulated and Constitutive Secretory Vesicles from the trans‐Golgi Network

Wang¹,

Thiele

Huttner

2000

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127

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“…Quatacker (1979), however, has shown that both GERL and terminal membranes of the Golgi complex stain with phosphotungstic acid and, therefore, these membranes may be rich in glycom-oteins. Following passage through-the Golgi complex and terminal glycosylation (Geissler et al, 1977), chromogranins Fig. 18.…”

Section: A 6 1 O F I H E O P O S S U M I N Daysmentioning

confidence: 99%

“…where they are glycosylated (Geissler et al, 1977). Membrane-bound enzymes, such as dopamine P-hydroxylase, would be simultaneously synthesized and processed by the GERL.…”

Section: A 6 1 O F I H E O P O S S U M I N Daysmentioning

confidence: 99%

Opossum adrenal medulla: II. Differentiation of the chromaffin cell

et al. 1987

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The ultrastructure of the opossum adrenal medulla was examined in its postnatal development. Maturation of chromaffin cells and genesis of chromaffin vesicles were of particular interest. The primitive sympathetic cell was seen to contain few organelles with no apparent polarity. Initial pheochromoblasts contained more organelles with some polarity. Endoplasmic reticulum and the Golgi complex increased as the pheochromoblasts matured, which suggested increased synthetic activity. Structures resembling Golgi/endoplasmic reticulum/lysosome (GERL) systems were seen in the pheochromoblasts. It is suggested that some of the components of the chromaffin vesicle may be processed by the GERL while others come directly through the Golgi complex. It is stressed that the developing pheochromoblast in the opossum presents an interesting model in which to study the genesis of the chromaffin vesicle.

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“…When analyzed by SDS-PAGE, peak 4 showed a predominance of a band of M, 72,000, with several faster migrating species also immunoreactive with anti-CGA antibodies. The natural CGA protein of M, 72,000 is known to be phosphorylated on serine residues (Settleman et al, 1985a), and is also glycosylated (Smith & Winkler, 1967;Geissler et al, 1977;Fischer-Colbrie et al, 1982). Therefore, the slightly later elution time of the recombinant CGA is expected.…”

Section: Verification Of the Sequence Of Recombinant Cgamentioning

confidence: 99%

“…CGA also binds calcium and catecholamines and self associates in the presence of calcium at high protein concentration (Cohn et al, 1981;Reiffen & Gratzl, 1986a,b;Leiser & Sherwood, 1989;Yo0 & Albanesi, 1990; Videen et al, 1992). It is difficult to obtain sufficient CGA of a single well-characterized molecular species for either structural or functional studies, because the protein is extensively proteolytically processed (Wohlfarter et al, 1988) and occurs in phosphorylated (Settleman et al, 1985a), glycosylated (Smith & Winkler, 1967;Geissler et al, 1977;Fischer-Colbrie et al, 1982), and proteoglycan forms (Falkensammer et al, 1985;Rosa et al, 1985;Gowda et al, 1990) as well. In order to pursue further studies with a defined CGA polypeptide, we have expressed the cDNA for bovine CGA in the pET3a vector system devised by Studier (Studier & Moffat, 1986;Studier et al, 1990).…”

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Effects of calcium on recombinant bovine chromogranin A

et al. 1992

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Bovine chromogranin A, the acidic calcium-binding protein characteristic of endocrine secretory vesicles, has been expressed in Escherichia coli using the pET3a vector system under T7 polymerase control. The expressed protein is located in the bacterial cytosol and can be purified from bacterial proteins by a heat treatment step, followed by gel filtration, anion-exchange, and reversed-phase chromatography. The purified recombinant chromogranin A has an apparent M, of ca. 72,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, in spite of its 432-amino acid polypeptide chain, consistent with observations on natural chromogranin A. The primary structure has been confirmed by mass spectral analysis of tryptic peptides, by Edman degradation of the intact protein, and by immunoreactivity with sequence-specific antibodies. Analysis by circular dichroism spectroscopy shows pH-and concentration-dependent spectra. The spectra are Ca2+-dependent from 5 to 40 P.M.

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Composition and biogenesis of complex carbohydrates of ox adrenal chromaffin granules

Cited by 64 publications

References 40 publications

Cholesterol is Required for the Formation of Regulated and Constitutive Secretory Vesicles from the trans‐Golgi Network

Cholesterol is Required for the Formation of Regulated and Constitutive Secretory Vesicles from the trans‐Golgi Network

Opossum adrenal medulla: II. Differentiation of the chromaffin cell

Effects of calcium on recombinant bovine chromogranin A

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