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Liu, K.-C., Pace, H., Larsson, E., Hossain, S., Kabedev, A., Shukla, A., . . . Lundmark, R. (2022). Membrane insertion mechanism of the caveola coat protein Cavin1. Proceedings of the National Academy of Sciences of the United States of America, 119(25), Article ID 2202295119.
Öppna denna publikation i ny flik eller fönster >>Membrane insertion mechanism of the caveola coat protein Cavin1
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2022 (Engelska)Ingår i: Proceedings of the National Academy of Sciences of the United States of America, ISSN 0027-8424, E-ISSN 1091-6490, Vol. 119, nr 25, artikel-id 2202295119Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Caveolae are small plasma membrane invaginations, important for control of membrane tension, signaling cascades, and lipid sorting. The caveola coat protein Cavin1 is essential for shaping such high curvature membrane structures. Yet, a mechanistic understanding of how Cavin1 assembles at the membrane interface is lacking. Here, we used model membranes combined with biophysical dissection and computational modeling to show that Cavin1 inserts into membranes. We establish that initial phosphatidylinositol (4, 5) bisphosphate [PI(4,5)P2]-dependent membrane adsorption of the trimeric helical region 1 (HR1) of Cavin1 mediates the subsequent partial separation and membrane insertion of the individual helices. Insertion kinetics of HR1 is further enhanced by the presence of flanking negatively charged disordered regions, which was found important for the coassembly of Cavin1 with Caveolin1 in living cells. We propose that this intricate mechanism potentiates membrane curvature generation and facilitates dynamic rounds of assembly and disassembly of Cavin1 at the membrane.

Ort, förlag, år, upplaga, sidor
Proceedings of the National Academy of Sciences, 2022
Nyckelord
caveolae, Cavin1, membrane curvature, membrane-shaping protein, protein-lipid interactions
Nationell ämneskategori
Biokemi och molekylärbiologi
Identifikatorer
urn:nbn:se:umu:diva-203198 (URN)10.1073/pnas.2202295119 (DOI)000838706900008 ()2-s2.0-85133725056 (Scopus ID)
Forskningsfinansiär
Vetenskapsrådet, 2018-05973Europeiska kommissionenKempestiftelsernaCancerfondenWallenbergstiftelserna
Tillgänglig från: 2023-01-18 Skapad: 2023-01-18 Senast uppdaterad: 2023-01-18Bibliografiskt granskad
Wang, T., Sarwar, M., Whitchurch, J. B., Collins, H. M., Green, T., Semenas, J., . . . Persson, J. L. (2022). PIP5K1α is Required for Promoting Tumor Progression in Castration-Resistant Prostate Cancer. Frontiers in Cell and Developmental Biology, 10, Article ID 798590.
Öppna denna publikation i ny flik eller fönster >>PIP5K1α is Required for Promoting Tumor Progression in Castration-Resistant Prostate Cancer
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2022 (Engelska)Ingår i: Frontiers in Cell and Developmental Biology, E-ISSN 2296-634X, Vol. 10, artikel-id 798590Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

PIP5K1α has emerged as a promising drug target for the treatment of castration-resistant prostate cancer (CRPC), as it acts upstream of the PI3K/AKT signaling pathway to promote prostate cancer (PCa) growth, survival and invasion. However, little is known of the molecular actions of PIP5K1α in this process. Here, we show that siRNA-mediated knockdown of PIP5K1α and blockade of PIP5K1α action using its small molecule inhibitor ISA-2011B suppress growth and invasion of CRPC cells. We demonstrate that targeted deletion of the N-terminal domain of PIP5K1α in CRPC cells results in reduced growth and migratory ability of cancer cells. Further, the xenograft tumors lacking the N-terminal domain of PIP5K1α exhibited reduced tumor growth and aggressiveness in xenograft mice as compared to that of controls. The N-terminal domain of PIP5K1α is required for regulation of mRNA expression and protein stability of PIP5K1α. This suggests that the expression and oncogenic activity of PIP5K1α are in part dependent on its N-terminal domain. We further show that PIP5K1α acts as an upstream regulator of the androgen receptor (AR) and AR target genes including CDK1 and MMP9 that are key factors promoting growth, survival and invasion of PCa cells. ISA-2011B exhibited a significant inhibitory effect on AR target genes including CDK1 and MMP9 in CRPC cells with wild-type PIP5K1α and in CRPC cells lacking the N-terminal domain of PIP5K1α. These results indicate that the growth of PIP5K1α-dependent tumors is in part dependent on the integrity of the N-terminal sequence of this kinase. Our study identifies a novel functional mechanism involving PIP5K1α, confirming that PIP5K1α is an intriguing target for cancer treatment, especially for treatment of CRPC.

Ort, förlag, år, upplaga, sidor
Frontiers Media S.A., 2022
Nyckelord
androgen receptor (AR), castration-resistant prostate cancer (CRPC), cyclin-dependent kinase (CDK), matrix metalloproteinases 9 (MMP9) PIP5K1α, phosphatidylinositol 4-phosphate 5 kinase (PIP5K1α), targeted therapy
Nationell ämneskategori
Cancer och onkologi
Forskningsämne
onkologi
Identifikatorer
urn:nbn:se:umu:diva-193614 (URN)10.3389/fcell.2022.798590 (DOI)000780059400001 ()35386201 (PubMedID)2-s2.0-85128078251 (Scopus ID)
Forskningsfinansiär
EU, Horisont 2020, 721297Barncancerfonden, TJ2015-0097Cancerfonden, CAN-2017-381Vetenskapsrådet, 2019-01318Stiftelsen för internationalisering av högre utbildning och forskning (STINT), IG2013-5595KempestiftelsernaCancerforskningsfonden i Norrland
Tillgänglig från: 2022-04-07 Skapad: 2022-04-07 Senast uppdaterad: 2023-05-23Bibliografiskt granskad
Larsson, E., Hubert, M. & Lundmark, R. (2020). Analysis of protein and lipid interactions using liposome co-sedimentation assays. In: Cedric M. Blouin (Ed.), Caveolae: methods and protocols (pp. 119-127). Humana Press
Öppna denna publikation i ny flik eller fönster >>Analysis of protein and lipid interactions using liposome co-sedimentation assays
2020 (Engelska)Ingår i: Caveolae: methods and protocols / [ed] Cedric M. Blouin, Humana Press, 2020, , s. 9s. 119-127Kapitel i bok, del av antologi (Refereegranskat)
Abstract [en]

The dynamic assembly of proteins at the membrane interphase is key to many cell biological processes such as the generation and stabilization of caveolae at the cell surface via coat proteins. The liposome co-sedimentation assay has been widely used for studies of protein and lipid interactions and has provided important information about binding mechanisms, lipid-binding specificity, and curvature preference of proteins. Here, we describe this technique in detail and how it can be used as a tool to address the membrane-binding ability and lipid specificity of caveolae-associated proteins.

Ort, förlag, år, upplaga, sidor
Humana Press, 2020. s. 9
Serie
Methods in Molecular Biology, ISSN 1064-3745, E-ISSN 1940-6029 ; 2169
Nyckelord
Caveolae coat, Cavin, Co-sedimentation, EHD2, Lipid specificity, Liposome pull down, Liposomes, Protein and lipid interactions, SUVs
Nationell ämneskategori
Biofysik Cell- och molekylärbiologi
Identifikatorer
urn:nbn:se:umu:diva-197951 (URN)10.1007/978-1-0716-0732-9_11 (DOI)2-s2.0-85086686139 (Scopus ID)978-1-0716-0731-2 (ISBN)978-1-0716-0734-3 (ISBN)978-1-0716-0732-9 (ISBN)
Tillgänglig från: 2022-07-08 Skapad: 2022-07-08 Senast uppdaterad: 2023-03-24Bibliografiskt granskad
Hubert, M., Larsson, E. & Lundmark, R. (2020). Keeping in touch with the membrane; protein- and lipid-mediated confinement of caveolae to the cell surface. Biochemical Society Transactions, 48, 155-163
Öppna denna publikation i ny flik eller fönster >>Keeping in touch with the membrane; protein- and lipid-mediated confinement of caveolae to the cell surface
2020 (Engelska)Ingår i: Biochemical Society Transactions, ISSN 0300-5127, E-ISSN 1470-8752, Vol. 48, s. 155-163Artikel, forskningsöversikt (Refereegranskat) Published
Abstract [en]

Caveolae are small Omega-shaped invaginations of the plasma membrane that play important roles in mechanosensing, lipid homeostasis and signaling. Their typical morphology is characterized by a membrane funnel connecting a spherical bulb to the membrane. Membrane funnels (commonly known as necks and pores) are frequently observed as transient states during fusion and fission of membrane vesicles in cells. However, caveolae display atypical dynamics where the membrane funnel can be stabilized over an extended period of time, resulting in cell surface constrained caveolae. In addition, caveolae are also known to undergo flattening as well as short-range cycles of fission and fusion with the membrane, requiring that the membrane funnel closes or opens up, respectively. This mini-review considers the transition between these different states and highlights the role of the protein and lipid components that have been identified to control the balance between surface association and release of caveolae.

Ort, förlag, år, upplaga, sidor
PORTLAND PRESS LTD, 2020
Nyckelord
caveolae, caveolin, cholesterol, dynamics, EHD2, pacsin2
Nationell ämneskategori
Biokemi och molekylärbiologi
Identifikatorer
urn:nbn:se:umu:diva-169355 (URN)10.1042/BST20190386 (DOI)000518382800015 ()32049332 (PubMedID)2-s2.0-85081069553 (Scopus ID)
Tillgänglig från: 2020-04-07 Skapad: 2020-04-07 Senast uppdaterad: 2023-03-23Bibliografiskt granskad
Hubert, M., Larsson, E., Vegesna, N. V., Ahnlund, M., Johansson, A. I., Moodie, L. W. K. & Lundmark, R. (2020). Lipid accumulation controls the balance between surface connection and scission of caveolae. eLIFE, 9, Article ID e55038.
Öppna denna publikation i ny flik eller fönster >>Lipid accumulation controls the balance between surface connection and scission of caveolae
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2020 (Engelska)Ingår i: eLIFE, E-ISSN 2050-084X, Vol. 9, artikel-id e55038Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Caveolae are bulb-shaped invaginations of the plasma membrane (PM) that undergo scission and fusion at the cell surface and are enriched in specific lipids. However, the influence of lipid composition on caveolae surface stability is not well described or understood. Accordingly, we inserted specific lipids into the cell PM via membrane fusion and studied their acute effects on caveolae dynamics. We demonstrate that sphingomyelin stabilizes caveolae to the cell surface, whereas cholesterol and glycosphingolipids drive caveolae scission from the PM. Although all three lipids accumulated specifically in caveolae, cholesterol and sphingomyelin were actively sequestered, whereas glycosphingolipids diffused freely. The ATPase EHD2 restricts lipid diffusion and counteracts lipid-induced scission. We propose that specific lipid accumulation in caveolae generates an intrinsically unstable domain prone to scission if not restrained by EHD2 at the caveolae neck. This work provides a mechanistic link between caveolae and their ability to sense the PM lipid composition.

Ort, förlag, år, upplaga, sidor
eLife Sciences Publications Ltd, 2020
Nationell ämneskategori
Biokemi och molekylärbiologi Cell- och molekylärbiologi
Identifikatorer
urn:nbn:se:umu:diva-172503 (URN)10.7554/eLife.55038 (DOI)000537207600001 ()32364496 (PubMedID)2-s2.0-85084964804 (Scopus ID)
Forskningsfinansiär
Vetenskapsrådet, 2017-04028Cancerfonden, CAN 2017/735Cancerfonden, CAN2014/746Kempestiftelserna
Tillgänglig från: 2020-07-02 Skapad: 2020-07-02 Senast uppdaterad: 2020-07-02Bibliografiskt granskad
Rodrigues, L., Schneider, F., Zhang, X., Larsson, E., Moodie, L. W. K., Dietz, H., . . . Hubert, M. (2019). Cellular uptake of self-assembled phytantriol-based hexosomes is independent of major endocytic machineries. Journal of Colloid and Interface Science, 553, 820-833
Öppna denna publikation i ny flik eller fönster >>Cellular uptake of self-assembled phytantriol-based hexosomes is independent of major endocytic machineries
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2019 (Engelska)Ingår i: Journal of Colloid and Interface Science, ISSN 0021-9797, E-ISSN 1095-7103, Vol. 553, s. 820-833Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Despite increasing interests in non-lamellar liquid crystalline dispersions, such as hexosomes, for drug delivery, little is known about their interactions with cells and mechanism of cell entry. Here we examine the cellular uptake of hexosomes based on phytantriol and mannide monooleate by HeLa cells using live cell microscopy in comparison to conventional liposomes. To investigate the importance of specific endocytosis pathways upon particle internalization, we silenced regulatory proteins of major endocytosis pathways using short interfering RNA. While endocytosis plays a significant role in liposome internalization, hexosomes are not taken up via endocytosis but through a mechanism that is dependent on cell membrane tension. Biophysical studies using biomembrane models highlighted that hexosomes have a high affinity for membranes and an ability to disrupt lipid layers. Our data suggest that direct biomechanical interactions of hexosomes with membrane lipids play a crucial role and that the unique morphology of hexosomes is vital for their membrane activity. Based on these results, we propose a mechanism, where hexosomes destabilize the bilayer, allowing them to "phase through" the membrane. Understanding parameters that influence the uptake of hexosomes is critical to establish them as carrier systems that can potentially deliver therapeutics efficiently to intracellular sites of action.

Ort, förlag, år, upplaga, sidor
Elsevier, 2019
Nyckelord
Hexosomes, Phytantriol, Mannide monooleate, Self-assembly, Nanostructure, Endocytosis, Cell take, Biomembrane models
Nationell ämneskategori
Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)
Identifikatorer
urn:nbn:se:umu:diva-164503 (URN)10.1016/j.jcis.2019.06.045 (DOI)000483454400086 ()31284226 (PubMedID)2-s2.0-85068359801 (Scopus ID)
Tillgänglig från: 2019-11-28 Skapad: 2019-11-28 Senast uppdaterad: 2023-03-23Bibliografiskt granskad
Moodie, L. W. K., Hubert, M., Zhou, X., Albers, M. F., Lundmark, R., Wanrooij, S. & Hedberg, C. (2019). Photoactivated Colibactin Probes Induce Cellular DNA Damage. Angewandte Chemie International Edition, 58(5), 1417-1421
Öppna denna publikation i ny flik eller fönster >>Photoactivated Colibactin Probes Induce Cellular DNA Damage
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2019 (Engelska)Ingår i: Angewandte Chemie International Edition, ISSN 1433-7851, E-ISSN 1521-3773, Vol. 58, nr 5, s. 1417-1421Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Colibactin is a small molecule produced by certain bacterial species of the human microbiota that harbour the pks genomic island. Pks(+) bacteria induce a genotoxic phenotype in eukaryotic cells and have been linked with colorectal cancer progression. Colibactin is produced in a benign, prodrug form which, prior to export, is enzymatically matured by the producing bacteria to its active form. Although the complete structure of colibactin has not been determined, key structural features have been described including an electrophilic cyclopropane motif, which is believed to alkylate DNA. To investigate the influence of the putative "warhead" and the prodrug strategy on genotoxicity, a series of photolabile colibactin probes were prepared that upon irradiation induced a pks(+) like phenotype in HeLa cells. Furthermore, results from DNA cross-linking and imaging studies of clickable analogues enforce the hypothesis that colibactin effects its genotoxicity by directly targeting DNA.

Nyckelord
click chemistry, colibactin, DNA damage, microbiome, photochemistry
Nationell ämneskategori
Biokemi och molekylärbiologi
Identifikatorer
urn:nbn:se:umu:diva-156892 (URN)10.1002/anie.201812326 (DOI)000458826100026 ()30506956 (PubMedID)2-s2.0-85059115203 (Scopus ID)
Tillgänglig från: 2019-03-11 Skapad: 2019-03-11 Senast uppdaterad: 2023-10-18Bibliografiskt granskad
Geh, K. J., Hubert, M. & Winter, G. (2018). Progress in formulation development and sterilisation of freeze-dried oligodeoxynucleotide-loaded gelatine nanoparticles. European journal of pharmaceutics and biopharmaceutics, 129, 10-20
Öppna denna publikation i ny flik eller fönster >>Progress in formulation development and sterilisation of freeze-dried oligodeoxynucleotide-loaded gelatine nanoparticles
2018 (Engelska)Ingår i: European journal of pharmaceutics and biopharmaceutics, ISSN 0939-6411, E-ISSN 1873-3441, Vol. 129, s. 10-20Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Oligodeoxynucleotide (ODN)-loaded gelatine nanoparticles (GNPs) have proven their outstanding potential in the treatment of allergic diseases such as equine asthma and canine atopic dermatitis, which are appropriate models for the corresponding human diseases. To encourage the development of a marketable product, long term stability and sterility needs to be ensured. In this work, we aimed to advance freeze-drying options to stabilise ODN-loaded GNPs. Matrix-assisted laser desorption/ionisation mass spectrometry time-of-flight was implemented as a versatile tool to assess ODN stability. With this method long-term storage stability of lyophilised ODN-loaded GNPs formulated in sucrose or trehalose was achieved. Controlled nucleation was further introduced to optimise the lyophilisation approach. This allowed shortening of the process in comparison to standard freeze-drying procedures. Particle sizes, polydispersity indices, ODN stability, residual moisture and glass transition temperature were maintained upon storage. Excipient portfolio was enlarged by novel amino acid containing formulations for lyophilisates. His emerged as an excellent excipient in stabilising lyophilised ODN-loaded GNPs, whereas addition of Arg and Gly revealed to be inadequate at accelerated conditions. Lastly, gamma irradiation was evaluated as a suitable sterilisation method of ODN-loaded GNPs.

Ort, förlag, år, upplaga, sidor
Elsevier, 2018
Nationell ämneskategori
Farmaceutiska vetenskaper
Identifikatorer
urn:nbn:se:umu:diva-150657 (URN)10.1016/j.ejpb.2018.05.016 (DOI)000438179500002 ()29777773 (PubMedID)2-s2.0-85047254615 (Scopus ID)
Tillgänglig från: 2018-08-28 Skapad: 2018-08-28 Senast uppdaterad: 2018-08-28Bibliografiskt granskad
Hubert, M., Larsson, E. & Lundmark, R. (2017). Caveolae dynamics is strongly influenced by the lipid composition of the plasma membrane. Paper presented at 19th IUPAB Congress / 11th EBSA Congress, JUL 16-20, 2017, British Biophys Soc, Edinburgh, SCOTLAND. European Biophysics Journal, 46, S121-S121
Öppna denna publikation i ny flik eller fönster >>Caveolae dynamics is strongly influenced by the lipid composition of the plasma membrane
2017 (Engelska)Ingår i: European Biophysics Journal, ISSN 0175-7571, E-ISSN 1432-1017, Vol. 46, s. S121-S121Artikel i tidskrift, Meeting abstract (Övrigt vetenskapligt) Published
Ort, förlag, år, upplaga, sidor
Springer, 2017
Nationell ämneskategori
Biofysik
Identifikatorer
urn:nbn:se:umu:diva-143671 (URN)000416406200233 ()
Konferens
19th IUPAB Congress / 11th EBSA Congress, JUL 16-20, 2017, British Biophys Soc, Edinburgh, SCOTLAND
Tillgänglig från: 2018-01-16 Skapad: 2018-01-16 Senast uppdaterad: 2018-06-09Bibliografiskt granskad
Holst, M. R., Vidal-Quadras, M., Larsson, E., Song, J., Hubert, M., Blomberg, J., . . . Lundmark, R. (2017). Clathrin-Independent Endocytosis Suppresses Cancer Cell Blebbing and Invasion. Cell Reports, 20(8), 1893-1905
Öppna denna publikation i ny flik eller fönster >>Clathrin-Independent Endocytosis Suppresses Cancer Cell Blebbing and Invasion
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2017 (Engelska)Ingår i: Cell Reports, E-ISSN 2211-1247, Vol. 20, nr 8, s. 1893-1905Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Cellular blebbing, caused by local alterations in cellsurface tension, has been shown to increase the invasiveness of cancer cells. However, the regulatory mechanisms balancing cell-surface dynamics and bleb formation remain elusive. Here, we show that an acute reduction in cell volume activates clathrinindependent endocytosis. Hence, a decrease in surface tension is buffered by the internalization of the plasma membrane (PM) lipid bilayer. Membrane invagination and endocytosis are driven by the tension- mediated recruitment of the membrane sculpting and GTPase-activating protein GRAF1 (GTPase regulator associated with focal adhesion kinase-1) to the PM. Disruption of this regulation by depleting cells of GRAF1 or mutating key phosphatidylinositol- interacting amino acids in the protein results in increased cellular blebbing and promotes the 3D motility of cancer cells. Our data support a role for clathrin-independent endocytic machinery in balancing membrane tension, which clarifies the previously reported role of GRAF1 as a tumor suppressor.

Ort, förlag, år, upplaga, sidor
Cell Press, 2017
Nyckelord
Endocytosis, clathrin-independent endocytosis, membrane blebbing, membrane tension, GRAF1, cancer invasion, cell migration, ARHGAP26, cell surface dynamics, GRAF1-dependent endocytosis
Nationell ämneskategori
Cell- och molekylärbiologi
Identifikatorer
urn:nbn:se:umu:diva-139144 (URN)10.1016/j.celrep.2017.08.006 (DOI)000408154300014 ()28834752 (PubMedID)2-s2.0-85028307075 (Scopus ID)
Tillgänglig från: 2017-09-15 Skapad: 2017-09-15 Senast uppdaterad: 2024-01-17Bibliografiskt granskad
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