Open Access Research article

Novel interactions of CLN5 support molecular networking between Neuronal Ceroid Lipofuscinosis proteins

Annina Lyly, Carina von Schantz, Claudia Heine, Mia-Lisa Schmiedt, Tessa Sipilä, Anu Jalanko and Aija Kyttälä*

Author Affiliations

National Institute for Health and Welfare (THL), Biomedicum Helsinki, Finland and FIMM, Institute for Molecular Medicine in Finland

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BMC Cell Biology 2009, 10:83  doi:10.1186/1471-2121-10-83

Published: 26 November 2009

Additional files

Additional file 1:

Interaction of CLN5 with endogenous NCL proteins in HeLa cells. The mouse Cln5-cDNA was expressed as a GST fusion protein and used for pull down analyses of endogenous NCL proteins from HeLa cell lysates. The bound proteins were immunoblotted and detected with specific antibodies: CLN2, rabbit polyclonal antibody 7951 [23], CLN3, polyclonal 385 antibody [21] PPT1, rabbit polyclonal antibody 8414 [19], and with anti-CLN6 antibody, which was a generous gift from Dr. S. Mole (London, UK) (Wheeler et al. Am J Hum Genet 70, 2002).

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Additional file 2:

Co-expression of the mutated CLN5 and wtPPT1 in HeLa cells. HeLa cells were transiently transfected with the mutated CLN5Fin (A) and wt CLN1/PPT1 (B). The cells were fixed with methanol 48 h post transfection, stained and analyzed by confocal microscopy. The Golgi complex is stained with GM130 (C). CLN5Fin and wt PPT co-localized only partially with the Golgi complex (D-F). Scale bar 10 μm.

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Additional file 3:

Facilitated lysosomal trafficking of the mutated CLN5 by PPT1 overexpression in SH-SY5Y cells. Human neuroblastoma cells (SH-SY5Y), were transiently transfected with wt CLN1/PPT1 and CLN5-Fin, carrying the most common vLINCL(Fin) causing mutation (A-E), or with CLN5-Fin alone (F-H). The cells were fixed with methanol 48 h post transfection, stained and analyzed by confocal microscopy. When CLN5-Fin was co-expressed with wt PPT1, CLN5-Fin was able to traffic to lysosomes with PPT1 (A-E). When CLN5-Fin was expressed alone, it retained in the ER (F-H). Scale bar 10 μm.

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