Induction of specific storage organelles by von Willebrand factor propolypeptide

Wagner, Denisa D.; Saffaripour, Simin; Bonfanti, Roberta; Sadler, J. Evan; Cramer, Elisabeth M.; Chapman, Barbara S.; Mayadas, Tanya N.

doi:10.1016/0092-8674(91)90648-i

Cited by 251 publications

(247 citation statements)

References 56 publications

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“…The observation of intracellular heterodimers, but only secretion of the recombinant MUC2 Cterminal homodimers, suggests that the MUC2 C-terminus does not contain sorting information targeting it into the regulated secretory pathway. This is in analogy with the vWF, where the C-terminus has no role in targeting the protein to the regulated secretory pathway [33]. The information for vWF targeting to the regulated secretory pathway has instead been localized to the Nterminal part of vWF.…”

Section: Discussionmentioning

confidence: 99%

The recombinant C-terminus of the human MUC2 mucin forms dimers in Chinese-hamster ovary cells and heterodimers with full-length MUC2 in LS 174T cells

Lidell

Johansson

Mörgelin

et al. 2003

Biochemical Journal

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The entire cDNA corresponding to the C-terminal cysteine-rich domain of the human MUC2 apomucin, after the serine-and threonine-rich tandem repeat, was expressed in Chinese-hamster ovary-K1 cells and in the human colon carcinoma cell line, LS 174T. The C-terminus was expressed as a fusion protein with the green fluorescent protein and mycTag sequences and the murine immunoglobulin κ-chain signal sequence to direct the protein to the secretory pathway. Pulse-chase studies showed a rapid conversion of the C-terminal monomer into a dimer in both Chinese-hamster ovary-K1 and LS 174T cells. Disulphide-bondstabilized dimers secreted into the media of both cell lines had a higher apparent molecular mass compared with the intracellular forms. The MUC2 C-terminus was purified from the spent culture medium and visualized by molecular electron microscopy. The dimer nature of the molecule was visible clearly and revealed that each monomer was attached to the other by a large globular domain. Gold-labelled antibodies against the mycTag or green fluorescent protein revealed that these were localized to the ends opposite to the parts responsible for the dimerization. The Cterminus expressed in LS 174T cells formed heterodimers with the full-length wild-type MUC2, but not with the MUC5AC mucin, normally expressed in LS 174T cells. The homodimers of the MUC2 C-termini were secreted continuously from the LS 174T cells, but no wild-type MUC2 secretion has been observed from these cells. This suggests that the information for sorting the MUC2 mucin into the regulated secretory pathway in cells having this ability is present in parts other than the C-terminus of MUC2.

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Section: Discussionmentioning

confidence: 99%

The recombinant C-terminus of the human MUC2 mucin forms dimers in Chinese-hamster ovary cells and heterodimers with full-length MUC2 in LS 174T cells

Lidell

Johansson

Mörgelin

et al. 2003

Biochemical Journal

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“…The affinity of regulated proteins for themselves and for others within the TGN would lead to intrinsic, albeit non-random, heterogeneity of aggregates formed in the TGN. This in turn could account for the diversity of secretory granule contents within the very same cell (see for example [153,154]), as well as the generation of morphologically distinct granule forms upon expression of a foreign protein such as von Willebrand factor (vWF) in AtT20 cells [155].…”

Section: The Example Of the Secretograninsmentioning

confidence: 99%

Sorting and processing of secretory proteins

Halban

Irminger

1994

Biochemical Journal

307

226

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“…However, one striking difference is the relative efficiencies of the sorting processes for soluble and membrane proteins. When transfected into AtT20 cells, only a small percentage of soluble proteins capable of storage in dense core granules are actually sorted to these organelles; most of the proteins are secreted constitutively (Burgess and Kelly, 1987;Wagner et al, 1991). The same inefficiency is some- Although the sorting information for P-selectin has been located to the distal 10-residue Cl and 17-residue C2 segments, the precise features required for sorting remain to be determined.…”

Section: "25i-labeled Monoclonal Antibody Binding To Cellsmentioning

confidence: 99%

Cytoplasmic domain of P-selectin (CD62) contains the signal for sorting into the regulated secretory pathway.

Disdier

Morrissey

Fugate

et al. 1992

MBoC

155

111

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P-selectin (CD62), formerly called GMP-140 or PADGEM, is a membrane protein located in secretory storage granules of platelets and endothelial cells. To study the mechanisms responsible for the targeting of P-selectin to storage granules, we transfected its cDNA into COS-7 and CHO-Ki cells, which lack a regulated exocytic pathway, or into AtT20 cells, which are capable of regulated secretion. P-selectin was expressed on the plasma membrane of COS-7 and CHO-Kl cells but was concentrated in storage granules of AtT20 cells. Immunogold electron microscopy indicated that the electron-dense granules containing P-selectin in AtT20 cells also stored the endogenous soluble hormone ACTH. Activation of AtT20 cells with 8-Br-cAMP increased the surface expression of P-selectin, consistent with agonist-induced fusion of granule membranes with the plasma membrane. Deletion of the last 23 amino acids of the 35-residue cytoplasmic domain resulted in delivery of Pselectin to the plasma membrane of AtT20 cells. Replacement of the cytoplasmic tail of tissue factor, a plasma membrane protein, with the cytoplasmic domain of P-selectin redirected the chimeric molecule to granules. We conclude that the cytoplasmic domain of P-selectin is both necessary and sufficient for sorting of membrane proteins into the regulated pathway of secretion.

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Induction of specific storage organelles by von Willebrand factor propolypeptide

Cited by 251 publications

References 56 publications

The recombinant C-terminus of the human MUC2 mucin forms dimers in Chinese-hamster ovary cells and heterodimers with full-length MUC2 in LS 174T cells

The recombinant C-terminus of the human MUC2 mucin forms dimers in Chinese-hamster ovary cells and heterodimers with full-length MUC2 in LS 174T cells

Sorting and processing of secretory proteins

Cytoplasmic domain of P-selectin (CD62) contains the signal for sorting into the regulated secretory pathway.

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