, Volume 62, Issue 5, pp 805–810 | Cite as

Cytoplasmic ends of tetraspanin 7 harbour epitopes recognised by autoantibodies in type 1 diabetes

  • Anne Eugster
  • Gloria Kraus
  • Vicky Lidzba
  • Denise Müller
  • Manja Jolink
  • Anette-G. Ziegler
  • Ezio BonifacioEmail author
Short Communication



The beta cell protein tetraspanin 7 is a target of autoantibodies in individuals with type 1 diabetes. The aim of this study was to identify autoantibody epitope-containing regions and key residues for autoantibody binding.


Autoantibody epitope regions were identified by immunoprecipitation of luciferase-tagged single or multiple tetraspanin 7 domains using tetraspanin 7 antibody-positive sera. Subsequently, amino acids (AAs) relevant for autoantibody binding were identified by single AA mutations.


In tetraspanin 7 antibody-positive sera, antibody binding was most frequent to tetraspanin 7 proteins that contained the NH2-terminal cytoplasmic domain 1 (C1; up to 39%) or COOH-terminal C3 (up to 22%). Binding to C3 was more frequent when the domain was expressed along with the flanking transmembrane domain, suggesting that conformation is likely to be important. Binding to external domains was not observed. Single AA mutations of C3 identified residues Y246, E247 and R239 as critical for COOH-terminal binding of 9/10, 10/10 and 8/10 sera tested, respectively. Mutation of cysteines adjacent to the transmembrane domain at either residues C235 or C236 resulted in both decreased (8/178 and 15/178 individuals, respectively; >twofold decrease) and increased (30/178 and 13/178 individuals, respectively; >twofold increase) binding in participant sera vs wild-type protein.


We hypothesise that conformation and, potentially, modification of protein terminal ends of tetraspanin 7 may be important for autoantibody binding in type 1 diabetes.


Autoantibodies Epitopes Glima 38 Tetraspanin 7 Type 1 diabetes mellitus 



Amino acid


Cytoplasmic domain (1/2/3)


Extracellular domain (1/2)


Relative light units


Standard deviation score


Transmembrane domain (1/2/3/4)




Contribution statement

EB and AGZ designed the study, contributed to the conduct of the study, the acquisition, analysis and interpretation of data, and drafted, reviewed and approved the manuscript. AE, GK, VL, DM and MJ contributed to the acquisition, analysis and interpretation of data, and drafted, reviewed and approved the manuscript. EB is the guarantor of this work.


This work was funded by the JDRF (2-SRA-2015-13-Q-R) and the DFG FZ 111. Funding agencies had no role in the design of the study or interpretation of the results.

Duality of interest

The authors declare that there is no duality of interest associated with the manuscript.


  1. 1.
    McLaughlin KA, Richardson CC, Ravishankar A et al (2016) Identification of tetraspanin-7 as a target of autoantibodies in type 1 diabetes. Diabetes 65(6):1690–1698. CrossRefPubMedGoogle Scholar
  2. 2.
    Passini N, Larigan JD, Genovese S et al (1995) The 37/40-kilodalton autoantigen in insulin-dependent diabetes mellitus is the putative tyrosine phosphatase IA-2. Proc Natl Acad Sci U S A 92(20):9412–9416. CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Okada Y, Taniguchi H, Shimada C (1976) High concentration of GABA and high glutamate decarboxylase activity in rat pancreatic islets and human insulinoma. Science 194(4265):620–622. CrossRefPubMedGoogle Scholar
  4. 4.
    Hemler ME (2005) Tetraspanin functions and associated microdomains. Nat Rev Mol Cell Biol 6(10):801–811. CrossRefPubMedGoogle Scholar
  5. 5.
    Thümer L, Adler K, Bonifacio E et al (2010) German new onset diabetes in the young incident cohort study: DiMelli study design and first-year results. Rev Diabet Stud 7:198–204. CrossRefGoogle Scholar
  6. 6.
    Ziegler A-G, Meier-Stiegen F, Winkler C, Bonifacio E (2012) Prospective evaluation of risk factors for the development of islet autoimmunity and type 1 diabetes during puberty –TEENDIAB: study design. Pediatr Diabetes 13(5):419–424. CrossRefPubMedGoogle Scholar
  7. 7.
    Walther D, Eugster A, Jergens S et al (2016) Tetraspanin 7 autoantibodies in type 1 diabetes. Diabetologia 59(9):1973–1976. CrossRefPubMedGoogle Scholar
  8. 8.
    Hall MP, Unch J, Binkowski BF et al (2012) Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chem Biol 7(11):1848–1857. CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Roll U, Turck CW, Gitelman SE et al (2000) Peptide mapping and characterisation of glycation patterns of the glima 38 antigen recognised by autoantibodies in type I diabetic patients. Diabetologia 43(5):598–608. CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Anne Eugster
    • 1
  • Gloria Kraus
    • 1
  • Vicky Lidzba
    • 1
  • Denise Müller
    • 1
  • Manja Jolink
    • 2
    • 3
  • Anette-G. Ziegler
    • 2
    • 3
    • 4
  • Ezio Bonifacio
    • 1
    • 5
    Email author
  1. 1.DFG-Centre for Regenerative Therapies Dresden, Faculty of Medicine Carl Gustav CarusTechnische Universität DresdenDresdenGermany
  2. 2.Institute of Diabetes ResearchHelmholtz Zentrum MünchenMunichGermany
  3. 3.Forschergruppe DiabetesTechnische Universität München, Klinikum Rechts der IsarMunichGermany
  4. 4.Forschergruppe Diabetes e.V. at Helmholtz Zentrum MünchenMunichGermany
  5. 5.Paul Langerhans Institute Dresden, Helmholtz Centre Munich, Faculty of Medicine Carl Gustav CarusTechnische Universität DresdenDresdenGermany

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