Atomic structures of fibrillar segments of hIAPP suggest tightly mated β-sheets are important for cytotoxicity

hIAPP fibrils are associated with Type-II Diabetes, but the link of hIAPP structure to islet cell death remains elusive. Here we observe that hIAPP fibrils are cytotoxic to cultured pancreatic β-cells, leading us to determine the structure and cytotoxicity of protein segments composing the amyloid spine of hIAPP. Using the cryoEM method MicroED, we discover that one segment, 19–29 S20G, forms pairs of β-sheets mated by a dry interface that share structural features with and are similarly cytotoxic to full-length hIAPP fibrils. In contrast, a second segment, 15–25 WT, forms non-toxic labile β-sheets. These segments possess different structures and cytotoxic effects, however, both can seed full-length hIAPP, and cause hIAPP to take on the cytotoxic and structural features of that segment. These results suggest that protein segment structures represent polymorphs of their parent protein and that segment 19–29 S20G may serve as a model for the toxic spine of hIAPP. DOI: http://dx.doi.org/10.7554/eLife.19273.001

[1]  T. Gonen,et al.  Atomic resolution structure determination by the cryo‐EM method MicroED , 2017, Protein science : a publication of the Protein Society.

[2]  Ewa A. Mirecka,et al.  β-Hairpin of Islet Amyloid Polypeptide Bound to an Aggregation Inhibitor , 2016, Scientific Reports.

[3]  Ewa A. Mirecka,et al.  Structural Characterization of Fibrils from Recombinant Human Islet Amyloid Polypeptide by Solid-State NMR: The Central FGAILS Segment Is Part of the β-Sheet Core , 2016, PloS one.

[4]  S. Kahn,et al.  The S20G substitution in hIAPP is more amyloidogenic and cytotoxic than wild-type hIAPP in mouse islets , 2016, Diabetologia.

[5]  K. Rhodes,et al.  The antibody aducanumab reduces Aβ plaques in Alzheimer’s disease , 2016, Nature.

[6]  D. Raleigh,et al.  Time-resolved studies define the nature of toxic IAPP intermediates, providing insight for anti-amyloidosis therapeutics , 2016, eLife.

[7]  T. Gonen,et al.  The collection of MicroED data for macromolecular crystallography , 2016, Nature Protocols.

[8]  T. P. Davis,et al.  Pancreatic β-Cell Membrane Fluidity and Toxicity Induced by Human Islet Amyloid Polypeptide Species , 2016, Scientific Reports.

[9]  D. Eisenberg,et al.  Crystal Structures of IAPP Amyloidogenic Segments Reveal a Novel Packing Motif of Out-of-Register Beta Sheets. , 2016, The journal of physical chemistry. B.

[10]  Nicholas K. Sauter,et al.  Structure of the toxic core of α-synuclein from invisible crystals , 2015, Nature.

[11]  Tamir Gonen,et al.  MicroED data collection and processing , 2015, Acta crystallographica. Section A, Foundations and advances.

[12]  Claudio Soto,et al.  Type 2 diabetes as a protein misfolding disease. , 2015, Trends in molecular medicine.

[13]  V. Yee,et al.  Crystal Structures of Polymorphic Prion Protein β1 Peptides Reveal Variable Steric Zipper Conformations. , 2015, Biochemistry.

[14]  G. Westermark,et al.  Heparan Sulfate Proteoglycans Are Important for Islet Amyloid Formation and Islet Amyloid Polypeptide-induced Apoptosis* , 2015, The Journal of Biological Chemistry.

[15]  Y. Miller,et al.  Orientations of residues along the β-arch of self-assembled amylin fibril-like structures lead to polymorphism. , 2015, Biomacromolecules.

[16]  S. Radford,et al.  Screening and classifying small molecule inhibitors of amyloid formation using ion mobility spectrometry-mass spectrometry , 2014, Nature Chemistry.

[17]  A. Miranker,et al.  Fiber-dependent and -independent toxicity of islet amyloid polypeptide. , 2014, Biophysical journal.

[18]  T. Gonen,et al.  Structure of catalase determined by MicroED , 2014, eLife.

[19]  Paul D. Adams,et al.  FEM: feature-enhanced map , 2014, Acta crystallographica. Section D, Biological crystallography.

[20]  Tamir Gonen,et al.  High-resolution structure determination by continuous rotation data collection in MicroED , 2014, Nature Methods.

[21]  G. Pappalardo,et al.  Molecular and cytotoxic properties of hIAPP17-29 and rIAPP17-29 fragments: a comparative study with the respective full-length parent polypeptides. , 2014, European journal of medicinal chemistry.

[22]  S. Kahn,et al.  Pathophysiology and treatment of type 2 diabetes: perspectives on the past, present, and future , 2014, The Lancet.

[23]  N. Amdursky,et al.  Apoptosis induced by islet amyloid polypeptide soluble oligomers is neutralized by diabetes-associated specific antibodies , 2014, Scientific Reports.

[24]  Tamir Gonen,et al.  Three-dimensional electron crystallography of protein microcrystals , 2013, eLife.

[25]  K. Diederichs,et al.  Better models by discarding data? , 2013, Acta crystallographica. Section D, Biological crystallography.

[26]  D. Eisenberg,et al.  Out-of-register β-sheets suggest a pathway to toxic amyloid aggregates , 2012, Proceedings of the National Academy of Sciences.

[27]  Ann Marie Schmidt,et al.  Sensitivity of amyloid formation by human islet amyloid polypeptide to mutations at residue 20. , 2012, Journal of molecular biology.

[28]  Mathias Jucker,et al.  The Amyloid State of Proteins in Human Diseases , 2012, Cell.

[29]  David Eisenberg,et al.  Atomic View of a Toxic Amyloid Small Oligomer , 2012, Science.

[30]  A. Miranker,et al.  Concentration‐dependent transitions govern the subcellular localization of islet amyloid polypeptide , 2012, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[31]  Ian S. Haworth,et al.  Fibril Structure of Human Islet Amyloid Polypeptide*♦ , 2011, The Journal of Biological Chemistry.

[32]  Jason T. Stevens,et al.  Structure-based design of non-natural amino-acid inhibitors of amyloid fibril formation , 2011, Nature.

[33]  Per Westermark,et al.  Islet amyloid polypeptide, islet amyloid, and diabetes mellitus. , 2011, Physiological reviews.

[34]  J. Esko,et al.  Heparan sulfate proteoglycans. , 2011, Cold Spring Harbor perspectives in biology.

[35]  S. Kahn,et al.  β-cell loss and β-cell apoptosis in human type 2 diabetes are related to islet amyloid deposition. , 2011, The American journal of pathology.

[36]  T. Sanke,et al.  Progressive deterioration of insulin secretion in Japanese type 2 diabetic patients in comparison with those who carry the S20G mutation of the islet amyloid polypeptide gene: A long‐term follow‐up study , 2011, Journal of diabetes investigation.

[37]  David Eisenberg,et al.  β2-microglobulin forms 3D domain-swapped amyloid fibrils with disulfide linkages , 2010, Nature Structural &Molecular Biology.

[38]  Fanling Meng,et al.  The flavanol (-)-epigallocatechin 3-gallate inhibits amyloid formation by islet amyloid polypeptide, disaggregates amyloid fibrils, and protects cultured cells against IAPP-induced toxicity. , 2010, Biochemistry.

[39]  Adrien Treuille,et al.  Predicting protein structures with a multiplayer online game , 2010, Nature.

[40]  R. L. Hull,et al.  Toxic oligomers and islet beta cell death: guilty by association or convicted by circumstantial evidence? , 2010, Diabetologia.

[41]  Vincent B. Chen,et al.  Correspondence e-mail: , 2000 .

[42]  L. Breydo,et al.  Fibrillar Oligomers Nucleate the Oligomerization of Monomeric Amyloid β but Do Not Seed Fibril Formation* , 2009, The Journal of Biological Chemistry.

[43]  David Eisenberg,et al.  Molecular basis for insulin fibril assembly , 2009, Proceedings of the National Academy of Sciences.

[44]  David Eisenberg,et al.  Molecular mechanisms for protein-encoded inheritance , 2009, Nature Structural &Molecular Biology.

[45]  David Eisenberg,et al.  Atomic structures of IAPP (amylin) fusions suggest a mechanism for fibrillation and the role of insulin in the process , 2009, Protein science : a publication of the Protein Society.

[46]  D. Otzen,et al.  Unique identification of supramolecular structures in amyloid fibrils by solid-state NMR spectroscopy. , 2009, Angewandte Chemie.

[47]  C. Ling,et al.  Mitochondrial dysfunction in pancreatic β-cells in Type 2 Diabetes , 2008, Molecular and Cellular Endocrinology.

[48]  S. Radford,et al.  Structural insights into the polymorphism of amyloid-like fibrils formed by region 20-29 of amylin revealed by solid-state NMR and X-ray fiber diffraction. , 2008, Journal of the American Chemical Society.

[49]  David Eisenberg,et al.  Atomic structure of the cross‐β spine of islet amyloid polypeptide (amylin) , 2008, Protein science : a publication of the Protein Society.

[50]  M. Köhler,et al.  Real-Time Monitoring of Apoptosis by Caspase-3-Like Protease Induced FRET Reduction Triggered by Amyloid Aggregation , 2008, Experimental diabetes research.

[51]  P. Butler,et al.  Islet amyloid in type 2 diabetes, and the toxic oligomer hypothesis. , 2008, Endocrine reviews.

[52]  Richard Leapman,et al.  Peptide conformation and supramolecular organization in amylin fibrils: constraints from solid-state NMR. , 2007, Biochemistry.

[53]  Elizabeth Head,et al.  Fibril specific, conformation dependent antibodies recognize a generic epitope common to amyloid fibrils and fibrillar oligomers that is absent in prefibrillar oligomers , 2007, Molecular Neurodegeneration.

[54]  R. Rizza,et al.  High Expression Rates of Human Islet Amyloid Polypeptide Induce Endoplasmic Reticulum Stress–Mediated β-Cell Apoptosis, a Characteristic of Humans With Type 2 but Not Type 1 Diabetes , 2007, Diabetes.

[55]  Heather T. McFarlane,et al.  Atomic structures of amyloid cross-β spines reveal varied steric zippers , 2007, Nature.

[56]  Chia-yu Lin,et al.  Toxic Human Islet Amyloid Polypeptide (h-IAPP) Oligomers Are Intracellular, and Vaccination to Induce Anti-Toxic Oligomer Antibodies Does Not Prevent h-IAPP–Induced β-Cell Apoptosis in h-IAPP Transgenic Mice , 2007, Diabetes.

[57]  Johannes D. Veldhuis,et al.  Human Islet Amyloid Polypeptide Oligomers Disrupt Cell Coupling, Induce Apoptosis, and Impair Insulin Secretion in Isolated Human Islets , 2007, Diabetes.

[58]  Airlie J. McCoy,et al.  Solving structures of protein complexes by molecular replacement with Phaser , 2006, Acta crystallographica. Section D, Biological crystallography.

[59]  Ralf Langen,et al.  Inhibition of human IAPP fibril formation does not prevent beta-cell death: evidence for distinct actions of oligomers and fibrils of human IAPP. , 2006, American journal of physiology. Endocrinology and metabolism.

[60]  S. Radford,et al.  The organization of aromatic side groups in an amyloid fibril probed by solid-state 2H and 19F NMR spectroscopy. , 2006, Journal of the American Chemical Society.

[61]  S. Kahn,et al.  Genetic background determines the extent of islet amyloid formation in human islet amyloid polypeptide transgenic mice. , 2005, American journal of physiology. Endocrinology and metabolism.

[62]  S. Kahn,et al.  Long-term treatment with rosiglitazone and metformin reduces the extent of, but does not prevent, islet amyloid deposition in mice expressing the gene for human islet amyloid polypeptide. , 2005, Diabetes.

[63]  Robert A. Grothe,et al.  Structure of the cross-β spine of amyloid-like fibrils , 2005, Nature.

[64]  Ueli Aebi,et al.  The parallel superpleated beta-structure as a model for amyloid fibrils of human amylin. , 2005, Journal of molecular biology.

[65]  J. Levy,et al.  Islet amyloid polypeptide gene promoter polymorphisms are not associated with Type 2 diabetes or with the severity of islet amyloidosis. , 2005, Biochimica et biophysica acta.

[66]  Randy J Read,et al.  Electronic Reprint Biological Crystallography Likelihood-enhanced Fast Translation Functions Biological Crystallography Likelihood-enhanced Fast Translation Functions , 2022 .

[67]  Kevin Cowtan,et al.  research papers Acta Crystallographica Section D Biological , 2005 .

[68]  Robert A. Rizza,et al.  β-Cell Deficit and Increased β-Cell Apoptosis in Humans With Type 2 Diabetes , 2003, Diabetes.

[69]  J. Critchley,et al.  The islet amyloid polypeptide (amylin) gene S20G mutation in Chinese subjects: Evidence for associations with type 2 diabetes and cholesterol levels , 2001, Clinical endocrinology.

[70]  D. Steiner,et al.  Islet Amyloid Development in a Mouse Strain Lacking Endogenous Islet Amyloid Polypeptide (IAPP) but Expressing Human IAPP , 2000, Molecular medicine.

[71]  N. Eberhardt,et al.  S20G mutant amylin exhibits increased in vitro amyloidogenicity and increased intracellular cytotoxicity compared to wild-type amylin. , 2000, The American journal of pathology.

[72]  B. Ahrén,et al.  Islet amyloid and type 2 diabetes mellitus. , 2000, The New England journal of medicine.

[73]  U Aebi,et al.  Amyloid fibril formation from full-length and fragments of amylin. , 2000, Journal of structural biology.

[74]  J. Bernhagen,et al.  Identification of a penta- and hexapeptide of islet amyloid polypeptide (IAPP) with amyloidogenic and cytotoxic properties. , 2000, Journal of molecular biology.

[75]  N. Eberhardt,et al.  Intracellular amyloidogenesis by human islet amyloid polypeptide induces apoptosis in COS-1 cells. , 1999, The American journal of pathology.

[76]  D. Raleigh,et al.  Effects of sequential proline substitutions on amyloid formation by human amylin20-29. , 1999, Biochemistry.

[77]  D. Schubert,et al.  Cytotoxic Amyloid Peptides Inhibit Cellular 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐Diphenyltetrazolium Bromide (MTT) Reduction by Enhancing MTT Formazan Exocytosis , 1997, Journal of neurochemistry.

[78]  L. Serpell,et al.  Common core structure of amyloid fibrils by synchrotron X-ray diffraction. , 1997, Journal of molecular biology.

[79]  D. Peterson,et al.  Mechanism of Cellular 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐Diphenyltetrazolium Bromide (MTT) Reduction , 1997, Journal of neurochemistry.

[80]  U Aebi,et al.  Polymorphic fibrillar assembly of human amylin. , 1997, Journal of structural biology.

[81]  T. Sanke,et al.  Missense Mutation of Amylin Gene (S20G) in Japanese NIDDM Patients , 1996, Diabetes.

[82]  W. Soeller,et al.  Spontaneous diabetes mellitus in transgenic mice expressing human islet amyloid polypeptide. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[83]  R. Palmiter,et al.  Islet amyloid formation associated with hyperglycemia in transgenic mice with pancreatic beta cell expression of human islet amyloid polypeptide. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[84]  N. Eberhardt,et al.  Human islet amyloid polypeptide expression in COS-1 cells. A model of intracellular amyloidogenesis. , 1995, The American journal of pathology.

[85]  C. Behl,et al.  Amyloid peptides are toxic via a common oxidative mechanism. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[86]  J. M. Griffiths,et al.  Rotational Resonance Solid-State NMR Elucidates a Structural Model of Pancreatic Amyloid , 1995 .

[87]  B. Yankner,et al.  Beta-amyloid neurotoxicity requires fibril formation and is inhibited by congo red. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[88]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.

[89]  H. Wolfson,et al.  Shape complementarity at protein–protein interfaces , 1994, Biopolymers.

[90]  Bruce A. Yankner,et al.  Pancreatic islet cell toxicity of amylin associated with type-2 diabetes mellitus , 1994, Nature.

[91]  C. Betsholtz,et al.  Islet amyloid polypeptide: pinpointing amino acid residues linked to amyloid fibril formation. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[92]  A J Day,et al.  Molecular and functional characterization of amylin, a peptide associated with type 2 diabetes mellitus. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[93]  D. Steiner,et al.  Conservation of the sequence of islet amyloid polypeptide in five mammals is consistent with its putative role as an islet hormone. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[94]  J. Rothbard,et al.  Amylin found in amyloid deposits in human type 2 diabetes mellitus may be a hormone that regulates glycogen metabolism in skeletal muscle. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[95]  D. W. Hayden,et al.  Amyloid fibrils in human insulinoma and islets of Langerhans of the diabetic cat are derived from a neuropeptide-like protein also present in normal islet cells. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[96]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[97]  M. L. Connolly.  Solvent-accessible surfaces of proteins and nucleic acids. , 1983, Science.

[98]  R. Doolittle,et al.  A simple method for displaying the hydropathic character of a protein. , 1982, Journal of molecular biology.

[99]  D. Longnecker,et al.  The relation of islet amyloid to the clinical type of diabetes. , 1981, Human pathology.

[100]  A. Gazdar,et al.  Continuous, clonal, insulin- and somatostatin-secreting cell lines established from a transplantable rat islet cell tumor. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[101]  F M Richards,et al.  Areas, volumes, packing and protein structure. , 1977, Annual review of biophysics and bioengineering.

[102]  B. Lee,et al.  The interpretation of protein structures: estimation of static accessibility. , 1971, Journal of molecular biology.

[103]  E. Opie.  THE RELATION OE DIABETES MELLITUS TO LESIONS OF THE PANCREAS. HYALINE DEGENERATION OF THE ISLANDS OE LANGERHANS , 1901, The Journal of experimental medicine.

[104]  David Eisenberg,et al.  Atomic structures of amyloid cross-beta spines reveal varied steric zippers. , 2007, Nature.

[105]  Robert A. Grothe,et al.  Structure of the cross-beta spine of amyloid-like fibrils. , 2005, Nature.

[106]  Robert A Rizza,et al.  Beta-cell deficit and increased beta-cell apoptosis in humans with type 2 diabetes. , 2003, Diabetes.

[107]  A. Kapurniotu,et al.  Amyloidogenicity and cytotoxicity of islet amyloid polypeptide. , 2001, Biopolymers.

[108]  M. Lutter,et al.  Biochemical pathways of caspase activation during apoptosis. , 1999, Annual review of cell and developmental biology.

[109]  P. Lansbury,et al.  The structural basis of pancreatic amyloid formation : isotope-edited spectroscopy in the solid state , 1992 .