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    • Intra-Golgi Vesicular Transport and the Secretory Pathway

    Klaus Fiedler

    In a line of research at the Memorial Sloan-Kettering Cancer Center (Prof. James Rothman`s laboratory) intra-Golgi transport (A) was analyzed in a model setting and the cytoplasmic domain of the putative cargo receptor p24-TMED2 was shown to interact with coatomer (the Golgi coat) suggestive of bimodal interaction of coatomer with different signals and bimodal sorting (B) . Today, numerous new elegant studies are suggestive of the tri- or multi-modal sorting capacity of coatomer and the structural analysis had proven the original assignment to coatomer subcomplexes that I had found. The transmembrane domain of TMED proteins contains an extended motif involving a glutamine residue close to the cytoplasmic domain which interacts with sphingomyelin 18 and permits transport of p24 from the endoplasmic reticulum (ER) and Golgi to later stations in the secretory route (see also Supplementary Data). Moreover, the p24 protein that I had analyzed (C), showed a distinct pattern of several signals within the transmembrane/cytoplasmic domains establishing the role of signals in forward and backward trafficking.
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    A  The Secretory Pathway  

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    B  Bimodal Interaction of Coatomer

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    C  p24/TMED Interactions          

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    D  Predicted Luminal Interactions

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    E  Wnt Interaction of the p24 Model

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    F  Carbohydrate Binding of Model

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    G  Collagen Binding in Docking

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    H  Collagen Binding of TMED10

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    I  Binding to X-Ray Structures?

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    J  Bacterial Protein Collagen Binding

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    K  Considering a Random Chain

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    L  Considering the Young's Modulus

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    M  Collagen Traffic: Solutions

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    N  X-Ray Structures TMED2/10

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    O  Docking: X-Ray Structures

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    P  Molecular Dynamics Approach

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    Q  Interaction Found in Strain

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    R  Morphing May Aid              

    Vesicular traffic and cargo sorting is thus intricately regulated and p24 has e.g. been recently shown to have proviral activity in dengue virus infection , and recently p23-TMED10 , another member of the TMED-family, was implicated in vaccinia virus infection . Lately, this sphingomyelin-binding motif, originally found in the p24-TMED2 and gamma secretase, has been described in several other polypeptides such as tetraspanin-7 . p23-TMED10 was previously found to interact with the presenilin complex, important in signaling in the Notch pathway and Alzheimer APP processing (D) . p24-TMED2 may interact with Wnt proteins similar to the recently discovered and confirmed interaction of mammalian Wnt3-Fz8, and, moreover, p23-TMED10 and p24-TMED2 may both interact with N-glycans and collagen as analyzed with molecular docking (E, F). Collagen interaction in docking is shown for modeled TMED2 and the solved TMED10 structure (G, H). Comparison of solved TMED2 and TMED10 structures with the PDB database and the structural overlay of TMED10 and the collagenase-domain of bacterial source including the molecular docking, validated the overall model-docking test (I, J). Surprisingly, in Drosophila it was found that logjam (p24/TMED2) was involved in collagen secretion in a genetic screen . Moreover, the interplay of TANGO1, the putative cargo receptor for (pro-)collagen, and TMED proteins has recently been found . In addition, in a recent study with a shTMED9 knockout in primary human colon cancer cells, it is found that among several coregulated expressed proteins, the Wnt11 protein and regulators of the Wnt-pathway are up- whereas, for example, the Anxa13 (see also the page on epithelia and polarity) and Col9a3 are significantly downregulated . With respect to collagen binding and vesicular transport a model is put forward that would assume that collagen flexibility before further assembly, allows incorporation into small carrier vesicles and not only megacarriers (K, L, M)(see also Talks). To further establish the carbohydrate interaction shown in (F) X-ray structures of both, TMED2 and TMED10 (N, O) were further scrutinized but led to little confirmation for TMED2. Thus a molecular dynamics approach was used that reaffirmingly allowed adequate visualization of hybrid N-glycan interaction with both, TMED10 and now TMED2 (P, Q). The hindrance in interaction due to crystallization seems surmounted, molecular morphing may aid in further visualization of modeled p24-TMED2 and structure poses (R). In view of transported cargo and the homo- and heterooligomeric nature of TMED proteins, it is tempting to speculate about a multitude and/or of bidentate interactions of receptor and ligand.

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    References
    1. Fiedler K, Veit M, Stamnes MA, Rothman JE. Bimodal interaction of coatomer with the p24 family of putative cargo receptors. Science. 1996;273:1396-9.
    2. Fiedler K, Rothman JE. Sorting determinants in the transmembrane domain of p24 proteins. J Biol Chem. 1997;272:24739-42.
    3. Dodonova SO, Diestelkoetter-Bachert P, von Appen A, Hagen WJH, Beck R, Beck M, et al. A structure of the COPI coat and the role of coat proteins in membrane vesicle assembly. Science. 2015;349(6244):195-8.
    4. Contreras F-X, Ernst AM, Haberkant P, Bjorkholm P, Lindahl E, Gonen B, et al. Molecular recognition of a single sphingolipid species by a protein`s transmembrane domain. Nature. 2012;481(7382):525-9.
    5. Zanini F, Pu S-Y, Bekerman E, Einav S, Quake SR. Single-cell transcriptional dynamics of flavivirus infection. Elife [Internet]. 2018;7:e32942.
    6. Sohn K, Orci L, Ravazzola M, Amherdt M, Bremser M, Lottspeich F, et al. A major transmembrane protein of Golgi-derived COPI-coated vesicles involved in coatomer binding. J Cell Biol. 1996;135(5):1239–1248.
    7. Luteijn RD, van Diemen F, Blomen VA, Boer IGJ, Manikam Sadasivam S, van Kuppevelt TH, et al. A genome-wide haploid genetic screen identifies heparan sulfate-associated genes and the macropinocytosis modulator TMED10 as factors supporting vaccinia virus infection. J Virol. 2019;93(13):e02160-18.
    8. Bjorkholm P, Ernst AM, Hacke M, Wieland F, Brugger B, von Heijne G. Identification of novel sphingolipid-binding motifs in mammalian membrane proteins. Biochim Biophys Acta. 2014;1838(8):2066-70.
    9. Chen F, Hasegawa H, Schmitt-Ulms G, Kawarai T, Bohm C, Katayama T, et al. TMP21 is a presenilin complex component that modulates gamma-secretase but not epsilon-secretase activity. Nature. 2006;440(7088):1208-12.
    10. Fiedler, K. The Wnt segment polarity pathway and TMED2 protein may interact via a lectin- and decoy-type mechanism. bioRxiv. 2016; https://doi.org/10.1101/056531.
    11. Ke H, Feng Z, Liu M, Sun T, Dai J, Ma M, Liu L-P, Ni J-Q, Pastor-Pareja JC. Collagen secretion screening in Drosophila supports a common secretory machinery and multiple Rab requirements. Journal of Genetics and Genomics. 2018; 45 299-313.
    12. Yang K, Feng Z, Pastor-Pareja JC. p24-Tango1 interactions ensure ER-Golgi interface stability and efficient transport. bioRxiv. 2024; https://doi.org/10.1101/2024.02.23.580604
    13. Mishra S, Bernal C, Silvano M, Anand S, Ruiz i Altaba A. The protein secretion modulator TMED9 drives CNIH4/TGFa/GLI signaling opposing TMED3-WNT-TCF to promote colon cancer metastases. Oncogene. 2019;38:5817–5837.

    Update 2019
    KEYWORDS for SlideShow caveolae caveolin VIP36 VIP21 flotillin Pfad Analyse pathway analysis analysed cavatellin reggie MVP17 VIP17 cytochrome b5 XIII V II mechanosignaling mechanosignalling mechanotransduction Mechanotransduktion tensegrity leguminous Bauhinia lectin Lektin plant-like CRD calcium-binding p24 TMED protein COP alpha beta gamma zeta epsilon coatomer conformer conformation stack cisternae secretory transport vesicle vesicular Vesikel secretion Sekretion brain cell Zelle fMRI voxel based morphometry VBM hypertensive crisis clinical decision modeling modelling modelling crises scaling Hypertensive Krisen blood pressure silica beads perfusion coating lung immuno-isolation Immunoisolation purification Blutdruck Bluthochdruck hypertension multivariate statistics regression neural network ranks dimensionality machine reduction endocytosis Endozytose transcytosis adsorptive Transzytose paracellular parazellulär transcellular transzellulär epithelia Epithelien apical basal basolateral lateral polarity Polarität ESEM environmental scanning endothelia endothelium Endothelien vasculature Gefässe blood pressure vessel lumen luminal MDCK COS SVEC 4-10 Eahy 926 ECV304 BAEC CPAE mass spectrometry gel electrophoresis SDS-PAGE 2-D affinity chromatography two-dimensional proteomics time of flight micro-sequencing confocal plasma membrane surface DIC detergent complex trans-Golgi apparatus plasmalemmal Apparat complex Komplex ER endoplasmatisches Retikulum endoplasmic reticulum Reduktase NAD(P)H desaturase nucleus Nukleus lipid sterol cholesterol glycosphingolipid Sphingolipide spingomyelin GM1 Gangliosid Ceramid ceramide Transporter Ionen-Kanal ion channel membrane curvature tension viscosity flow LDL temperature low-density lipoprotein high-density merocrine Merokrin paracrine signaling