Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Oct;173(4):1066-9.
doi: 10.1111/bjd.13868. Epub 2015 Aug 19.

Heterogeneity in the properties of mutant secreted lymphocyte antigen 6/urokinase receptor-related protein 1 (SLURP1) in Mal de Meleda

O Adeyo  1 , M Oberer  2 , M Ploug  3 , L G Fong  1 , S G Young  1   4 , A P Beigneux  1
Affiliations

Heterogeneity in the properties of mutant secreted lymphocyte antigen 6/urokinase receptor-related protein 1 (SLURP1) in Mal de Meleda

O Adeyo et al. Br J Dermatol. 2015 Oct.
No abstract available

PubMed Disclaimer

Figures

Fig 1
Fig 1
Schematic of SLURP1 based on the crystal structure of other Ly6 proteins. The homology model of human SLURP1 was generated with the protein fold recognition server Phyre 2 using the 3D structure of LYNX1 as the template. The protein (excluding the signal peptide) is visualized with the PyMOL Molecular Graphics System. SLURP1 contains 10 cysteines, and all are disulfide-bonded (C25 with C50, C28 with C37, C43 with C73, C77 with C93, and C94 with C99). The five disulfide bonds are depicted in yellow. The locations of all missense mutations causing mal de Meleda are noted; three involve conserved cysteines of the Ly6 domain. The protein, like all Ly6 proteins, forms a three-fingered structural motif. The mutants that were secreted in low amounts and which led to protein multimers were located in the core of the protein; the two mutants that yielded a significant amount of monomers (G86R and P82S) are located at the tip of the third finger of the three-fingered motif. C-term, carboxyl terminus; N-term, amino terminus.
Fig 2
Fig 2
Heterogeneity in the properties of SLURP1 mutants associated with mal de Meleda. All experiments shown were repeated twice with virtually identical results. All samples were denatured in 1% lithium dodecyl sulfate for 10 min at 70° C. Where indicated, samples were reduced in 50 mM dithiothreitol. For each expression system, actin was used as a loading control. (a) Western blots depicting the expression of untagged WT-SLURP1 and SLURP1 mutants in transfected CHO cells. Shown (from the top) is a western blot of medium under nonreducing conditions (media, NR), a western blot of medium under reducing conditions (media, R) and a western blot of the cell extracts under reducing conditions (cells, R). Western blots were performed with a polyclonal antibody against human SLURP1 (Novus). All mutants were expressed, but they exhibited different levels of secretion. The secretion of SLURP1-R71H, SLURP1-R71P, SLURP1-C77R, SLURP1-C94S, SLURP1-L98P, and SLURP1-C99Y was reduced by 83–95% when compared to WT-SLURP1. For WT-SLURP1, monomers (~11 kDa, arrowhead) accounted for 37% of secreted SLURP1; for SLURP1-G86R, 24%; for SLURP1-P82S, 13%; for other SLURP1 mutants, 0.3–2%. (b) Western blots depicting the expression of untagged WT-SLURP1 and SLURP1 mutants in transfected HaCaT cells (a human keratinocyte cell line). In HaCaT cells, the secretion of the R71H, R71P, C77R, C94S, L98P, and C99Y mutants were reduced by >97%; the effects of the G86R and P82S mutations were more modest, and monomers were easily detected. SLURP1-W15R (with a mutation in the signal peptide) was not expressed, consistent with findings with a myc-tagged SLURP1-W15R construct. (c) Western blots depicting the expression of WT and mutant SLURP1 proteins in Drosophila S2 cells. These proteins were tagged at the carboxyl-terminus with sequences encoding uPAR Ly6 domain III, and therefore had a higher molecular weight. Nonreduced fusion proteins were detected with a uPAR-specific monoclonal antibody (R24); reduced proteins were detected with the SLURP1 polyclonal antibody. In Drosophila S2 cells, the production and synthesis of the different SLURP1 constructs was variable. Monomers (~20 kDa, arrowhead) accounted for 81–92% of WT-SLURP1, SLURP1-G86R, and SLURP1-P82S in the medium, whereas they accounted only for 7–43% of the SLURP1 with the other mutants.
See this image and copyright information in PMC

References

    1. Favre B, Plantard L, Aeschbach L, et al. SLURP1 is a late marker of epidermal differentiation and is absent in mal de meleda. J Invest Dermatol. 2007;127:301–308. - PubMed
    1. Beigneux AP, Franssen R, Bensadoun A, et al. Chylomicronemia with a mutant GPIHBP1 (Q115P) that cannot bind lipoprotein lipase. Arterioscler Thromb Vasc Biol. 2009;29:956–962. - PMC - PubMed
    1. Franssen R, Young SG, Peelman F, et al. Chylomicronemia with low postheparin lipoprotein lipase levels in the setting of GPIHBP1 defects. Circ Cardiovasc Genet. 2010;3:169–178. - PMC - PubMed
    1. Olivecrona G, Ehrenborg E, Semb H, et al. Mutation of conserved cysteines in the ly6 domain of GPIHBP1 in familial chylomicronemia. J Lipid Res. 2010;51:1535–1545. - PMC - PubMed
    1. Plengpanich W, Young SG, Khovidhunkit W, et al. Multimerization of GPIHBP1 and familial chylomicronemia from a serine-to-cysteine substitution in gpihbp1's ly6 domain. J Biol Chem. 2014;289:19491–19499. - PMC - PubMed

MeSH terms