Congenital disorder of glycosylation caused by starting site-specific variant in syntaxin-5.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
28 10 2021
28 10 2021
Historique:
received:
10
04
2020
accepted:
05
10
2021
entrez:
29
10
2021
pubmed:
30
10
2021
medline:
31
12
2021
Statut:
epublish
Résumé
The SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) protein syntaxin-5 (Stx5) is essential for Golgi transport. In humans, the STX5 mRNA encodes two protein isoforms, Stx5 Long (Stx5L) from the first starting methionine and Stx5 Short (Stx5S) from an alternative starting methionine at position 55. In this study, we identify a human disorder caused by a single missense substitution in the second starting methionine (p.M55V), resulting in complete loss of the short isoform. Patients suffer from an early fatal multisystem disease, including severe liver disease, skeletal abnormalities and abnormal glycosylation. Primary human dermal fibroblasts isolated from these patients show defective glycosylation, altered Golgi morphology as measured by electron microscopy, mislocalization of glycosyltransferases, and compromised ER-Golgi trafficking. Measurements of cognate binding SNAREs, based on biotin-synchronizable forms of Stx5 (the RUSH system) and Förster resonance energy transfer (FRET), revealed that the short isoform of Stx5 is essential for intra-Golgi transport. Alternative starting codons of Stx5 are thus linked to human disease, demonstrating that the site of translation initiation is an important new layer of regulating protein trafficking.
Identifiants
pubmed: 34711829
doi: 10.1038/s41467-021-26534-y
pii: 10.1038/s41467-021-26534-y
pmc: PMC8553859
doi:
Substances chimiques
Protein Isoforms
0
Qa-SNARE Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
6227Informations de copyright
© 2021. The Author(s).
Références
Jahn, R. & Scheller, R. H. SNAREs — engines for membrane fusion. Nat. Rev. Mol. Cell Biol. 7, 631–631 (2006).
pubmed: 16912714
doi: 10.1038/nrm2002
Banfield, D. K., Lewis, M. J. & Pelham, H. R. B. A SNARE-like protein required for traffic through the Golgi complex. Nature 375, 806–809 (1995).
pubmed: 7596416
doi: 10.1038/375806a0
Parlati, F. et al. Topological restriction of SNARE-dependent membrane fusion. Nature 407, 194–198 (2000).
pubmed: 11001058
doi: 10.1038/35025076
Parlati, F. et al. Distinct SNARE complexes mediating membrane fusion in Golgi transport based on combinatorial specificity. Proc. Natl Acad. Sci. USA 99, 5424–5429 (2002).
pubmed: 11959998
pmcid: 122785
doi: 10.1073/pnas.082100899
Xu, Y., Martin, S., James, D. E. & Hong, W. GS15 forms a SNARE complex with syntaxin 5, GS28, and Ykt6 and is implicated in traffic in the early cisternae of the Golgi apparatus. Mol. Biol. Cell 13, 3493–3507 (2002).
pubmed: 12388752
pmcid: 129961
doi: 10.1091/mbc.e02-01-0004
Bentley, M. et al. SNARE status regulates tether recruitment and function in homotypic COPII vesicle fusion. J. Biol. Chem. 281, 38825–38833 (2006).
pubmed: 17038314
doi: 10.1074/jbc.M606044200
Dascher, C., Matteson, J. & Balch, W. E. Syntaxin 5 regulates endoplasmic reticulum to Golgi transport. J. Biol. Chem. 269, 29363–29366 (1994).
pubmed: 7961911
doi: 10.1016/S0021-9258(18)43884-7
Rowe, T., Dascher, C., Bannykh, S., Plutner, H. & Balch, W. E. Role of vesicle-associated syntaxin 5 in the assembly of pre-Golgi intermediates. Science 279, 696–700 (1998).
pubmed: 9445473
doi: 10.1126/science.279.5351.696
Xu, D., Joglekar, A. P., Williams, A. L. & Hay, J. C. Subunit structure of a mammalian ER/Golgi SNARE complex. J. Biol. Chem. 275, 39631–39639 (2000).
pubmed: 11035026
doi: 10.1074/jbc.M007684200
Hay, J. C. et al. Localization, dynamics, and protein interactions reveal distinct roles for ER and Golgi SNAREs. J. Cell Biol. 141, 1489–1502 (1998).
pubmed: 9647643
pmcid: 2133002
doi: 10.1083/jcb.141.7.1489
Paek, I. et al. ERS-24, a mammalian v-SNARE implicated in vesicle traffic between the ER and the Golgi. J. Cell Biol. 137, 1017–1028 (1997).
pubmed: 9166403
pmcid: 2136225
doi: 10.1083/jcb.137.5.1017
Zhang, T. et al. Ykt6 forms a SNARE complex with syntaxin 5, GS28, and Bet1 and participates in a late stage in endoplasmic reticulum-Golgi transport. J. Biol. Chem. 276, 27480–27487 (2001).
pubmed: 11323436
doi: 10.1074/jbc.M102786200
Linders, P. T., van der Horst, C., ter Beest, M. & van den Bogaart, G. Stx5-mediated ER–Golgi transport in mammals and yeast. Cells 8, 780 (2019).
pmcid: 6721632
doi: 10.3390/cells8080780
Malsam, J. & Söllner, T. H. Organization of SNAREs within the Golgi stack. Cold Spring Harb. Perspect. Biol. 3, a005249–a005249 (2011).
pubmed: 21768609
pmcid: 3179334
doi: 10.1101/cshperspect.a005249
Tai, G. et al. Participation of the syntaxin 5/Ykt6/GS28/GS15 SNARE complex in transport from the early/recycling endosome to the trans-Golgi network. Mol. Biol. Cell 15, 4011–4022 (2004).
pubmed: 15215310
pmcid: 515336
doi: 10.1091/mbc.e03-12-0876
Dickinson, M. E. et al. High-throughput discovery of novel developmental phenotypes. Nature 537, 508–514 (2016).
pubmed: 27626380
pmcid: 5295821
doi: 10.1038/nature19356
Koscielny, G. et al. The International Mouse Phenotyping Consortium Web Portal, a unified point of access for knockout mice and related phenotyping data. Nucleic Acids Res. 42, D802–D809 (2014).
pubmed: 24194600
doi: 10.1093/nar/gkt977
Hui, N. et al. An isoform of the Golgi t-SNARE, syntaxin 5, with an endoplasmic reticulum retrieval signal. Mol. Biol. Cell 8, 1777–1787 (1997).
pubmed: 9307973
pmcid: 305736
doi: 10.1091/mbc.8.9.1777
Gao, G. & Banfield, D. K. Multiple features within the syntaxin Sed5p mediate its Golgi localization. Traffic 21, 274–296 (2020).
pubmed: 31883188
doi: 10.1111/tra.12720
Dominguez, M. et al. gp25L/emp24/p24 protein family members of the cis-Golgi network bind both COP I and II coatomer. J. Cell Biol. 140, 751–765 (1998).
pubmed: 9472029
pmcid: 2141742
doi: 10.1083/jcb.140.4.751
Miyazaki, K. et al. Contribution of the long form of syntaxin 5 to the organization of the endoplasmic reticulum. J. Cell Sci. 125, 5658–5666 (2012).
pubmed: 23077182
doi: 10.1242/jcs.105304
Suga, K., Saito, A., Tomiyama, T., Mori, H. & Akagawa, K. The syntaxin 5 isoforms Syx5 and Syx5L have distinct effects on the processing of β-amyloid precursor protein. J. Biochem. 146, 905–915 (2009).
pubmed: 19720618
doi: 10.1093/jb/mvp138
Avci, D. et al. The intramembrane protease SPP impacts morphology of the endoplasmic reticulum by triggering degradation of morphogenic proteins. J. Biol. Chem. 294, 2786–2800 (2019).
pubmed: 30578301
doi: 10.1074/jbc.RA118.005642
Hay, J. C., Hirling, H. & Scheller, R. H. Mammalian vesicle trafficking proteins of the endoplasmic reticulum and Golgi apparatus. J. Biol. Chem. 271, 5671–5679 (1996).
pubmed: 8621431
doi: 10.1074/jbc.271.10.5671
Shestakova, A., Suvorova, E., Pavliv, O., Khaidakova, G. & Lupashin, V. Interaction of the conserved oligomeric Golgi complex with t-SNARE syntaxin5a/Sed5 enhances intra-Golgi SNARE complex stability. J. Cell Biol. 179, 1179–1192 (2007).
pubmed: 18086915
pmcid: 2140037
doi: 10.1083/jcb.200705145
Linders, P. T. A., Peters, E., ter Beest, M., Lefeber, D. J. & van den Bogaart, G. Sugary logistics gone wrong: membrane trafficking and congenital disorders of glycosylation. Int. J. Mol. Sci. 21, 4654 (2020).
pmcid: 7369703
doi: 10.3390/ijms21134654
Fung, C. W. et al. COG5-CDG with a mild neurohepatic presentation. JIMD Rep. 3, 67–70 (2012).
pubmed: 23430875
doi: 10.1007/8904_2011_61
Paesold-Burda, P. et al. Deficiency in COG5 causes a moderate form of congenital disorders of glycosylation. Hum. Mol. Genet. 18, 4350–4356 (2009).
pubmed: 19690088
doi: 10.1093/hmg/ddp389
Palmigiano, A. et al. MALDI-MS profiling of serum O-glycosylation and N-glycosylation in COG5-CDG. J. Mass Spectrom. 52, 372–377 (2017).
pubmed: 28444691
doi: 10.1002/jms.3936
Rymen, D. et al. COG5-CDG: expanding the clinical spectrum. Orphanet J. Rare Dis. 7, 94 (2012).
pubmed: 23228021
pmcid: 3697985
doi: 10.1186/1750-1172-7-94
Zhang, T. et al. The mammalian protein (rbet1) homologous to yeast Bet1p is primarily associated with the pre-Golgi intermediate compartment and is involved in vesicular transport from the endoplasmic reticulum to the Golgi apparatus. J. Cell Biol. 139, 1157–1168 (1997).
pubmed: 9382863
pmcid: 2140212
doi: 10.1083/jcb.139.5.1157
Mallard, F. et al. Early/recycling endosomes-to-TGN transport involves two SNARE complexes and a Rab6 isoform. J. Cell Biol. 156, 653–664 (2002).
pubmed: 11839770
pmcid: 2174079
doi: 10.1083/jcb.200110081
Dingjan, I. et al. Endosomal and phagosomal SNAREs. Physiol. Rev. 98, 1465–1492 (2018).
pubmed: 29790818
doi: 10.1152/physrev.00037.2017
Chiu, C.-F. et al. ZFPL1, a novel ring finger protein required for cis-Golgi integrity and efficient ER-to-Golgi transport. EMBO J. 27, 934–947 (2008).
pubmed: 18323775
pmcid: 2323254
doi: 10.1038/emboj.2008.40
Gleeson, P. A. et al. p230 is associated with vesicles budding from the trans-Golgi network. J. Cell Sci. 109, 2811–2821 (1996).
pubmed: 9013329
doi: 10.1242/jcs.109.12.2811
Jaiman, A. & Thattai, M. Golgi compartments enable controlled biomolecular assembly using promiscuous enzymes. eLife 9, e49573 (2020).
pubmed: 32597757
pmcid: 7360365
doi: 10.7554/eLife.49573
Reynders, E. et al. Golgi function and dysfunction in the first COG4-deficient CDG type II patient. Hum. Mol. Genet. 18, 3244–3256 (2009).
pubmed: 19494034
pmcid: 2722986
doi: 10.1093/hmg/ddp262
Climer, L. K., Pokrovskaya, I. D., Blackburn, J. B. & Lupashin, V. V. Membrane detachment is not essential for COG complex function. Mol. Biol. Cell 29, 964–974 (2018).
pubmed: 29467253
pmcid: 5896934
doi: 10.1091/mbc.E17-11-0694
Oka, T. et al. Genetic analysis of the subunit organization and function of the conserved oligomeric Golgi (COG) complex studies of COG5- and COG7-deficient mammalian cells. J. Biol. Chem. 280, 32736–32745 (2005).
pubmed: 16051600
doi: 10.1074/jbc.M505558200
Glick, B. S. & Nakano, A. Membrane traffic within the Golgi apparatus. Annu. Rev. Cell Dev. Biol. 25, 113–132 (2009).
pubmed: 19575639
pmcid: 2877624
doi: 10.1146/annurev.cellbio.24.110707.175421
Galea, G., Bexiga, M. G., Panarella, A., O’Neill, E. D. & Simpson, J. C. A high-content screening microscopy approach to dissect the role of Rab proteins in Golgi-to-ER retrograde trafficking. J. Cell Sci. 128, 2339–2349 (2015).
pubmed: 25999475
doi: 10.1242/jcs.167973
Lippincott-Schwartz, J., Roberts, T. H. & Hirschberg, K. Secretory protein trafficking and organelle dynamics in living cells. Annu. Rev. Cell Dev. Biol. 16, 557–589 (2000).
pubmed: 11031247
pmcid: 4781643
doi: 10.1146/annurev.cellbio.16.1.557
Boncompain, G. et al. Synchronization of secretory protein traffic in populations of cells. Nat. Methods 9, 493–493 (2012).
pubmed: 22406856
doi: 10.1038/nmeth.1928
Bindels, D. S. et al. mScarlet: a bright monomeric red fluorescent protein for cellular imaging. Nat. Methods 14, 53–56 (2017).
pubmed: 27869816
doi: 10.1038/nmeth.4074
Verboogen, D. R. J., González Mancha, N., ter Beest, M. & van den Bogaart, G. Fluorescence Lifetime Imaging Microscopy reveals rerouting of SNARE trafficking driving dendritic cell activation. eLife 6, e23525 (2017).
Griesbeck, O., Baird, G. S., Campbell, R. E., Zacharias, D. A. & Tsien, R. Y. Reducing the environmental sensitivity of yellow fluorescent protein mechanism and applications. J. Biol. Chem. 276, 29188–29194 (2001).
pubmed: 11387331
doi: 10.1074/jbc.M102815200
Casey, J. R., Grinstein, S. & Orlowski, J. Sensors and regulators of intracellular pH. Nat. Rev. Mol. Cell Biol. 11, 50–61 (2010).
pubmed: 19997129
doi: 10.1038/nrm2820
Antonin, W., Holroyd, C., Tikkanen, R., Höning, S. & Jahn, R. The R-SNARE endobrevin/VAMP-8 mediates homotypic fusion of early endosomes and late endosomes. Mol. Biol. Cell 11, 3289–3298 (2000).
pubmed: 11029036
pmcid: 14992
doi: 10.1091/mbc.11.10.3289
Bajno, L. et al. Focal exocytosis of Vamp3-containing vesicles at sites of phagosome formation. J. Cell Biol. 149, 697–706 (2000).
pubmed: 10791982
pmcid: 2174839
doi: 10.1083/jcb.149.3.697
Hong, W. SNAREs and traffic. Biochim. Biophys. Acta 1744, 120–144 (2005).
pubmed: 15893389
doi: 10.1016/j.bbamcr.2005.03.014
Manderson, A. P., Kay, J. G., Hammond, L. A., Brown, D. L. & Stow, J. L. Subcompartments of the macrophage recycling endosome direct the differential secretion of IL-6 and TNFα. J. Cell Biol. 178, 57–69 (2007).
pubmed: 17606866
pmcid: 2064421
doi: 10.1083/jcb.200612131
Murray, R. Z. A role for the phagosome in cytokine secretion. Science 310, 1492–1495 (2005).
pubmed: 16282525
doi: 10.1126/science.1120225
Amberger, J. S., Bocchini, C. A., Schiettecatte, F., Scott, A. F. & Hamosh, A. OMIM.org: Online Mendelian Inheritance in Man (OMIM
pubmed: 25428349
doi: 10.1093/nar/gku1205
Pedersen, A. G. & Nielsen, H. Neural network prediction of translation initiation sites in eukaryotes: perspectives for EST and genome analysis. Proc. Int Conf. Intell. Syst. Mol. Biol. 5, 226–233 (1997).
pubmed: 9322041
Kochetov, A. V. Alternative translation start sites and hidden coding potential of eukaryotic mRNAs. BioEssays 30, 683–691 (2008).
pubmed: 18536038
doi: 10.1002/bies.20771
Kozak, M. Regulation of translation via mRNA structure in prokaryotes and eukaryotes. Gene 361, 13–37 (2005).
pubmed: 16213112
doi: 10.1016/j.gene.2005.06.037
Oka, T., Ungar, D., Hughson, F. M. & Krieger, M. The COG and COPI Complexes Interact to Control the Abundance of GEARs, a Subset of Golgi Integral Membrane Proteins. Mol. Biol. Cell 15, 2423–2435 (2004).
pubmed: 15004235
pmcid: 404034
doi: 10.1091/mbc.e03-09-0699
Ohtsubo, K. & Marth, J. D. Glycosylation in cellular mechanisms of health and disease. Cell 126, 855–867 (2006).
pubmed: 16959566
doi: 10.1016/j.cell.2006.08.019
Freeze, H. H., Chong, J. X., Bamshad, M. J. & Ng, B. G. Solving glycosylation disorders: fundamental approaches reveal complicated pathways. Am. J. Hum. Genet. 94, 161–175 (2014).
pubmed: 24507773
pmcid: 3928651
doi: 10.1016/j.ajhg.2013.10.024
Fisher, P. & Ungar, D. Bridging the gap between glycosylation and vesicle traffic. Front. Cell Dev. Biol. 4, 15 (2016).
Blackburn, J. B., Kudlyk, T., Pokrovskaya, I. & Lupashin, V. V. More than just sugars: conserved oligomeric Golgi complex deficiency causes glycosylation-independent cellular defects. Traffic 19, 463–480 (2018).
pubmed: 29573151
pmcid: 5948163
doi: 10.1111/tra.12564
Foulquier, F. et al. Conserved oligomeric Golgi complex subunit 1 deficiency reveals a previously uncharacterized congenital disorder of glycosylation type II. Proc. Natl Acad. Sci. USA 103, 3764–3769 (2006).
pubmed: 16537452
pmcid: 1450151
doi: 10.1073/pnas.0507685103
Foulquier, F. et al. A new inborn error of glycosylation due to a Cog8 deficiency reveals a critical role for the Cog1–Cog8 interaction in COG complex formation. Hum. Mol. Genet. 16, 717–730 (2007).
pubmed: 17220172
doi: 10.1093/hmg/ddl476
Kranz, C. et al. COG8 deficiency causes new congenital disorder of glycosylation type IIh. Hum. Mol. Genet 16, 731–741 (2007).
pubmed: 17331980
doi: 10.1093/hmg/ddm028
Miller, V. J. & Ungar, D. Re’COG’nition at the Golgi. Traffic 13, 891–897 (2012).
pubmed: 22300173
doi: 10.1111/j.1600-0854.2012.01338.x
Morava, E. et al. A common mutation in the COG7 gene with a consistent phenotype including microcephaly, adducted thumbs, growth retardation, VSD and episodes of hyperthermia. Eur. J. Hum. Genet. 15, 638–645 (2007).
pubmed: 17356545
doi: 10.1038/sj.ejhg.5201813
Ng, B. G. et al. Molecular and clinical characterization of a Moroccan Cog7 deficient patient. Mol. Genet. Metab. 91, 201–204 (2007).
pubmed: 17395513
pmcid: 1941618
doi: 10.1016/j.ymgme.2007.02.011
Wu, X. et al. Mutation of the COG complex subunit gene COG7 causes a lethal congenital disorder. Nat. Med. 10, 518–523 (2004).
pubmed: 15107842
doi: 10.1038/nm1041
Hucthagowder, V. et al. Loss-of-function mutations in ATP6V0A2 impair vesicular trafficking, tropoelastin secretion and cell survival. Hum. Mol. Genet. 18, 2149–2165 (2009).
pubmed: 19321599
pmcid: 2685755
doi: 10.1093/hmg/ddp148
Jansen, J. C. et al. CCDC115 deficiency causes a disorder of Golgi homeostasis with abnormal protein glycosylation. Am. J. Hum. Genet. 98, 310–321 (2016).
pubmed: 26833332
pmcid: 4746332
doi: 10.1016/j.ajhg.2015.12.010
Jansen, J. C. et al. TMEM199 deficiency is a disorder of Golgi homeostasis characterized by elevated aminotransferases, alkaline phosphatase, and cholesterol and abnormal glycosylation. Am. J. Hum. Genet. 98, 322–330 (2016).
pubmed: 26833330
pmcid: 4746368
doi: 10.1016/j.ajhg.2015.12.011
Jansen, E. J. R. et al. ATP6AP1 deficiency causes an immunodeficiency with hepatopathy, cognitive impairment and abnormal protein glycosylation. Nat. Commun. 7, 11600–11600 (2016).
pubmed: 27231034
pmcid: 4894975
doi: 10.1038/ncomms11600
Foulquier, F. et al. TMEM165 deficiency causes a congenital disorder of glycosylation. Am. J. Hum. Genet. 91, 15–26 (2012).
pubmed: 22683087
pmcid: 3397274
doi: 10.1016/j.ajhg.2012.05.002
Ashikov, A. et al. Integrating glycomics and genomics uncovers SLC10A7 as essential factor for bone mineralization by regulating post-Golgi protein transport and glycosylation. Hum. Mol. Genet. 27, 3029–3045 (2018).
pubmed: 29878199
doi: 10.1093/hmg/ddy213
Park, J. H. et al. SLC39A8 deficiency: a disorder of manganese transport and glycosylation. Am. J. Hum. Genet. 97, 894–903 (2015).
pubmed: 26637979
pmcid: 4678430
doi: 10.1016/j.ajhg.2015.11.003
Witkos, T. M. et al. GORAB scaffolds COPI at the trans-Golgi for efficient enzyme recycling and correct protein glycosylation. Nat. Commun. 10, 1–18 (2019).
doi: 10.1038/s41467-018-08044-6
Zhong, W. Golgi during development. Cold Spring Harb. Perspect. Biol. 3, a005363 (2011).
Zhao, H. Membrane trafficking in osteoblasts and osteoclasts: new avenues for understanding and treating skeletal diseases. Traffic 13, 1307–1314 (2012).
pubmed: 22759194
pmcid: 3705567
doi: 10.1111/j.1600-0854.2012.01395.x
Wagner, T., Dieckmann, M., Jaeger, S., Weggen, S. & Pietrzik, C. U. Stx5 is a novel interactor of VLDL-R to affect its intracellular trafficking and processing. Exp. Cell Res. 319, 1956–1972 (2013).
pubmed: 23701949
doi: 10.1016/j.yexcr.2013.05.010
Bogaert, A., Fernandez, E. & Gevaert, K. N-terminal proteoforms in human disease. Trends Biochem. Sci. 45, 308–320 (2020).
pubmed: 32001092
doi: 10.1016/j.tibs.2019.12.009
Morelle, W. & Michalski, J.-C. Analysis of protein glycosylation by mass spectrometry. Nat. Protoc. 2, 1585–1602 (2007).
pubmed: 17585300
doi: 10.1038/nprot.2007.227
van Scherpenzeel, M., Steenbergen, G., Morava, E., Wevers, R. A. & Lefeber, D. J. High-resolution mass spectrometry glycoprofiling of intact transferrin for diagnosis and subtype identification in the congenital disorders of glycosylation. Transl. Res. 166, 639–649.e1 (2015).
pubmed: 26307094
doi: 10.1016/j.trsl.2015.07.005
Vissers, L. E. L. M. et al. A de novo paradigm for mental retardation. Nat. Genet. 42, 1109–1112 (2010).
pubmed: 21076407
doi: 10.1038/ng.712
Nikopoulos, K. et al. Next-generation sequencing of a 40 Mb linkage interval reveals TSPAN12 mutations in patients with familial exudative vitreoretinopathy. Am. J. Hum. Genet. 86, 240–247 (2010).
pubmed: 20159111
pmcid: 2820179
doi: 10.1016/j.ajhg.2009.12.016
Jolanda, I., de Vries, M., Adema, G. J., Punt, C. J. A. & Figdor, C. G. Phenotypical and functional characterization of clinical-grade dendritic cells. In Adoptive Immunotherapy: Methods and Protocols (eds. Ludewig, B. & Hoffmann, M. W.) 113–125 (Humana Press, 2005).
Ran, F. A. et al. Genome engineering using the CRISPR-Cas9 system. Nat. Protoc. 8, 2281–2308 (2013).
pubmed: 24157548
pmcid: 3969860
doi: 10.1038/nprot.2013.143
Virtanen, P. et al. SciPy 1.0: fundamental algorithms for scientific computing in Python. Nat. Methods 17, 261–272 (2020).
pubmed: 32015543
pmcid: 7056644
doi: 10.1038/s41592-019-0686-2
Schägger, H. Tricine–SDS-PAGE. Nat. Protoc. 1, 16–22 (2006).
pubmed: 17406207
doi: 10.1038/nprot.2006.4
Wickham, H. ggplot2: Elegant Graphics for Data Analysis. (Springer-Verlag New York, 2016).