Two complimentary sets of reagents were employed: one that inhibited IAPP secretion, but maintained the level of production of IAPP and a second that increased IAPP secretion but did not increase the amount of IAPP produced. Deamidation has also been shown to promote amyloid formation by otherwise nonamyloidogenic peptide fragments of hIAPP [48]. Green, C. Goldsbury, T. Mini et al., “Full-length rat amylin forms fibrils following substitution of single residues from human amylin,”, A. Abedini, F. L. Meng, and D. P. Raleigh, “A single-point mutation converts the highly amyloidogenic human islet amyloid polypeptide into a potent fibrillization inhibitor,”, F. Meng, D. P. Raleigh, and A. Abedini, “Combination of kinetically selected inhibitors in trans leads to highly effective inhibition of amyloid formation,”, L.-M. Yan, M. Tatarek-Nossol, A. Velkova, A. Kazantzis, and A. Kapurniotu, “Design of a mimic of nonamyloidogenic and bioactive human islet amyloid polypeptide (IAPP) as nanomolar affinity inhibitor of IAPP cytotoxic fibrillogenesis,”, S. Luca, W.-M. Yau, R. Leapman, and R. Tycko, “Peptide conformation and supramolecular organization in amylin fibrils: constraints from solid-state NMR,”, J. J. W. Wiltzius, S. A. Sievers, M. R. Sawaya et al., “Atomic structure of the cross-, A. T. Alexandrescu, “Amide proton solvent protection in amylin fibrils probed by quenched hydrogen exchange NMR,”, L. Wang, C. T. Middleton, S. Singh et al., “2DIR spectroscopy of human amylin fibrils reflects stable, D. Radovan, V. Smirnovas, and R. Winter, “Effect of pressure on islet amyloid polypeptide aggregation: revealing the polymorphic nature of the fibrillation process,”, V. Wineman-Fisher, Y. Atsmon-Raz, and Y. Miller, “Orientations of residues along the, R. Tycko, “Physical and structural basis for polymorphism in amyloid fibrils,”, C. S. Goldsbury, G. J. S. Cooper, K. N. Goldie et al., “Polymorphic fibrillar assembly of human amylin,”, S. Bedrood, Y. Li, J. M. Isas et al., “Fibril structure of human islet amyloid polypeptide,”, L. E. Buchanan, E. B. Dunkelberger, H. Q. Tran et al., “Mechanism of IAPP amyloid fibril formation involves an intermediate with a transient, T. M. Doran, E. A. Anderson, S. E. Latchney, L. A. Opanashuk, and B. L. Nilsson, “Turn nucleation perturbs amyloid, A. Abedini, R. Gupta, P. Marek et al., “Role of posttranslational modifications in amyloid formation,” in, E. B. Dunkelberger, L. E. Buchanan, P. Marek, P. Cao, D. P. Raleigh, and M. T. Zanni, “Deamidation accelerates amyloid formation and alters amylin fiber structure,”, M. R. Nilsson, M. Driscoll, and D. P. Raleigh, “Low levels of asparagine deamidation can have a dramatic effect on aggregation of amyloidogenic peptides: implications for the study of amyloid formation,”, S. Sakagashira, T. Sanke, T. Hanabusa et al., “Missense mutation of amylin gene (S20G) in Japanese NIDDM patients,”, S. Sakagashira, H. J. Hiddinga, K. Tateishi et al., “S20G mutant amylin exhibits increased, Z. Ma, G. T. Westermark, S. Sakagashira et al., “Enhanced in vitro production of amyloid-like fibrils from mutant (S20G) islet amyloid polypeptide,”, P. Cao, L.-H. Tu, A. Abedini et al., “Sensitivity of amyloid formation by human islet amyloid polypeptide to mutations at residue 20,”, A. Born, M. Havenith, C. Herrmann, and S. Ebbinghaus, “Protein stabilization by macromolecular crowding through enthalpy rather than entropy,”, M. Sarkar, A. E. Smith, and G. J. Pielak, “Impact of reconstituted cytosol on protein stability,”, M. D. Shtilerman, T. T. Ding, and P. T. Lansbury Jr., “Molecular crowding accelerates fibrillization of, L. A. Munishkina, E. M. Cooper, V. N. Uversky, and A. L. Fink, “The effect of macromolecular crowding on protein aggregation and amyloid fibril formation,”, Q. Ma, J.-B. hIAPP1–19 binds near the surface, similar to rat IAPP1–19, at acidic pH when His-18 is protonated, indicating that the net charge of residue 18 is important in controlling the orientation [93, 95]. e role of ER stress in, ER stress was not detected in studies of cultured islets that, and produce active caspase-. Porcine IAPP is far less amyloidogenic than hIAPP and the prevention of amyloid formation by transplantation of porcine islets prolongs islet graft survival [126]. IAPP is stored with insulin, in the granule and is released in response to the stimuli that, In this review we discuss the physical chemical properties, F : Primary sequences of human CGRP and of IAPP fro, in italics and underlined. The authors proposed that the EPR structure could represent an alternative polymorph. e American Journal of Physiology: Cell Physiology, on IAPP aggregation in the presence of an isola, cells in type diabetes: toxic islet amyloid, -cell dysfunction: the emerging connection, Proceedings of the National Academy of Sciences. e, studied in detail and seems ripe for further investigation. hIAPP can transiently adopt α-helix and β-strand conformations that could be important intermediate species on the fibrillization pathway. The results are consistent with an extracellular origin of islet amyloid. The biologically active sequences all contain a disulfide bridge between Cys-2 and Cys-7 and have an amidated C-terminus. II diabetes. Early studies, involving Ala scanning of short peptides derived from IAPP, supported this conjecture [57, 58]. -cell dysfunction and death in the disease and to the failure of islet transplants. The CT receptor has two splice variants and there are 3 relevant RAMPs, so there are six different subtypes of IAPP receptors. ApoE mouse knockou, that includes the N-terminal prosequence, denoted here by, ing to negatively charged membranes, negatively charged, heparanase in a double transgenic mouse model that over-, e factors that trigger islet amyloid formation, of perlecan [, , ]. The literature on IAPP mutations has been critically reviewed in 2013 and, in the interest of space, we refer the interested reader to that work for a more detailed discussion [12]. –, . The reason for the significant enhancement in the rate of amyloid formation by the Ser-20 to Gly mutation of hIAPP is still unknown. Quantitative mutational studies of amyloid fibril stability and of the kinetics of amyloid formation are much more challenging than studies with soluble, monomeric, globular proteins. Residual TFA from HPLC purification can affect amyloid formation and issues with lot to lot variability of ostensibly pure commercial IAPP have been reported [56]. 6. cholesterol have also been examined []. The initiation site for islet amyloid formation in vivo is controversial and there are conflicting reports in the literature. Finally, we propose that two different self-assembled fibril-like forms of amylin can interact to form a new fibril-like amylin. Diabetes-associated amyloidosis is not associated with visible recruitment of macrophages for removal of amyloid or islet debris. Islet amyloid has been detected in transplanted human islets in a patient that suffered islet graft failure and has been shown to form rapidly after transplantation of human islets into nude mice [4, 123–125]. IAPP Is Synthesized as a Preprohormone, []. Here we review what computer, in vitro, in vivo, and pharmacological experiments tell us about the accumulation and deposition of the oligomers of the (Aβ, tau), α-synuclein, IAPP, and superoxide dismutase 1 proteins, which have been the mainstream concept underlying Alzheimer's disease (AD), Parkinson's disease (PD), type II diabetes (T2D), and amyotrophic lateral sclerosis (ALS) research, respectively, for many years. This is of interest because Ser-28 and Ser-29 are located at the interface of the two symmetrically related columns of hIAPP monomers in the fibril structure and are involved in networks of hydrogen bonded interactions (Figure 3). (b) Primary sequence of the 67-residue proform of human IAPP. , no. I, in controlling gastric emptying, and in the supp, glucagon release [–]. The Leu side chain has approximately the same size and shape as Asn, but cannot hydrogen bond and is nonpolar. dene a precise, three-dimensional, high resolution structure. All rights reserved. However other studies with rodent models in which IAPP is overexpressed are consistent with an intracellular origin [4, 114]. In addition, it is important to bear in mind that amyloid fibrils are polymorphic and thus the alternative structures could well represent different polymorphs [39–42]. The polypeptide is positively charged at and below physiological pH with a net charge ranging from 2 to 4 depending upon the pKa’s of the N-terminus and His-18 and the pH. Interestingly, it has been noted that the smaller oligomers formed by IDPs are more toxic to cells than their larger aggregates. e Leu side chain has appr, the data than would experiments involving less co, nongenetically coded amino acids. The data presented here also propitiate new therapy opportunities, whether creating more effective drugs to diabetes or implementing a specific treatment to the patients with these mutations. The two peptides have reasonable sequence similarity but diverge the most within the segment corresponding to residues 20 and 29 [19]. Ester to Amide Switch Peptides Provide a Simple Method for Preparing Monomeric Islet Amyloid Polypeptide under Physiologically Relevant Conditions and Facilitate Investigations of Amyloid Formation. The effects of crowding agents and osmolytes on amyloid formation by IAPP are reviewed in detail in this volume by Gao and Winter [169]. We report here that at cytotoxic concentrations, amylin forms voltage-dependent, relatively nonselective, ion-permeable channels However other studies wi, some transgenic mouse models have high copy numbers of, used a cultured transgenic islet model to show that secretion, increased IAPP secretion but did not increase the amoun, extracellular origin of islet amyloid. These are important considerations since high percentages of anionic lipids significantly promote IAPP membrane interactions and because gangliosides and cholesterol modulate hIAPP membrane interactions [111, 148]. Because amylin also became the name of a pharmaceutical company, we have preferred IAPP. There is also interest in combining leptin and hIAPP for the treatment of obesity [175]. In addition, Ser-20 is located in the loop/bend region between the two β-strands in all of the models of hIAPP amyloid fibrils. NMR studies of a nonphysiological variant of hIAPP with a free C-terminal carboxyl group provided evidence for intermolecular interactions involving His-18 and Tyr-37 at pH 5.5 and it was suggested that these interactions play a role in the early stages of amyloid formation by hIAPP. However, despite the important role of chaperones on protein folding, less is known about chaperonal modulation of protein aggregation and fibrillation considering different classes of chaperones. An interesting alternative model, which differs from both the NMR and the fragment based model, has been proposed based upon EPR studies conducted with a set spin labeled variants of hIAPP incorporated into the polypeptide via Cys mutations. It is worth bearing in mind that some transgenic mouse models have high copy numbers of the human IAPP gene and can produce high levels of hIAPP. Furthermore, the T2DM-mediated increase in AD risk during aging might be attributed to an interaction of amylin with AD-related APs through evolvability, in which amylin protofibrillar formation presumably caused by adiponectin (APN) resistance could increase protofibril formation of AD-related APs in evolvability and subsequently lead to T2DM promotion of AD through antagonistic pleiotropy in aging. M, substitution of residues , , and i, corresponding amino acids of hIAPP led to a weakly amy-, loidogenic polypeptide even though it still contained the . Amylin is an endocrine hormone peptide that consists of 37 residues and is the main component of extracellular amyloid deposits found in the pancreas of most type 2 diabetes patients. (a) Primary sequence of human IAPP. Experimental challenges include relating reductionist biophysical experiments to the situation in vivo and understanding the connection between mouse models that highly overexpress hIAPP and human β-cell physiology. Analysis of the His-18 Gln mutant revealed that the rate of IAPP amyloid formation was still pH dependent between pH 5 and 8, thereby showing that the charge state of the N-terminus is an important factor modulating the rate of amyloid formation, even though the N-terminal region of IAPP is not part of the core β-sheet structure. The cellular environment determines the structure and function of proteins. in vivo Time resolved 2DIR studies provide a possible resolution of the apparent conundrum [44]. can be found in other reviews in this issue. The possible role of IAPP aggregation in the complications of diabetes has yet to be fully defined and the potential role of IAPP in type-1 diabetes remains to be elucidated [16–18]. Given the very different data used to construct the models, it is striking and reassuring that they share many common features. A growing body of evidence suggests that β-cell failure in type 2 diabetes correlates with the formation of pancreatic islet amyloid deposits, indicating that islet amyloid may have an important role in β-cell loss in this disease. There is no correlation between the presence of SAP and islet amyloid deposition. Each column contains two in-register parallel β-sheets. Calculations performed at the level of the linearized Poisson-Boltzmann (PB) approach show that Arg-11 makes significant unfavorable electrostatic interactions but that His-18 does not do so when its side chain is neutral. (c) View rotated by 90° showing the arrangement of the two stacks as a ribbon diagram. In the solid state NMR model, the N-terminal strand encompasses residues 8 to 17 and the C-terminal strand residues 28 to 37. IAPP is synthesized as a 89-residue preproform (Figure 3) [7]. ere are no clinically a, inhibitors of IAPP toxicity and very few, if any. Islet amyloid polypeptide listed as IAPP. The final product depends upon how the ring is opened. Song, and D. P. Raleigh, “Amyloid formation by pro-islet amyloid polypeptide processing intermediates: examination of the role of protein heparan sulfate interactions and implications for islet amyloid formation in type 2 diabetes,”, H. Wang and D. P. Raleigh, “The ability of insulin to inhibit the formation of amyloid by pro-islet amyloid polypeptide processing intermediates is significantly reduced in the presence of sulfated glycosaminoglycans,”, E. T. A. S. Jaikaran and A. Clark, “Islet amyloid and type 2 diabetes: from molecular misfolding to islet pathophysiology,”, R. L. Hull, G. T. Westermark, P. Westermark, and S. E. Kahn, “Islet amyloid: a critical entity in the pathogenesis of type 2 diabetes,”, J. C. Hutton, “The internal pH and membrane potential of the insulin-secretory granule,”, S. B. P. Charge, E. J. P. De Koning, and A. Clark, “Effect of pH and insulin on fibrillogenesis of islet amyloid polypeptide in vitro,”, A. Abedini and D. P. Raleigh, “The role of His-18 in amyloid formation by human islet amyloid polypeptide,”, P. Westermark, Z.-C. Li, G. T. Westermark, A. Leckström, and D. F. Steiner, “Effects of beta cell granule components on human islet amyloid polypeptide fibril formation,”, S. Janciauskiene, S. Eriksson, E. Carlemalm, and B. Ahrén, “B cell granule peptides affect human islet amyloid polypeptide (IAPP) fibril formation in vitro,”, E. T. A. S. Jaikaran, M. R. Nilsson, and A. Clark, “Pancreatic beta-cell granule peptides form heteromolecular complexes which inhibit islet amyloid polypeptide fibril formation,”, J. L. Larson and A. D. Miranker, “The mechanism of insulin action on islet amyloid polypeptide fiber formation,”, J. D. Knight, J. cell mass are key features of TD [, ]. Fox, T. Snollaerts, C. Errecart Casanova, A. Calciano, L. A. Nogaj, and D. A. Moffet, “Selection for nonamyloidogenic mutants of islet amyloid polypeptide (IAPP) identifies an extended region for amyloidogenicity,”, P. Marek, A. Abedini, B. in vitro Last, E. Rhoades, and A. D. Miranker, “Islet amyloid polypeptide demonstrates a persistent capacity to disrupt membrane integrity,”, J. R. Brender, E. L. Lee, M. A. Cavitt, A. Gafni, D. G. Steel, and A. Ramamoorthy, “Amyloid fiber formation and membrane disruption are separate processes localized in two distinct regions of IAPP, the type-2-diabetes-related peptide,”, M. F. M. Engel, L. Khemtémourian, C. C. Kleijer et al., “Membrane damage by human islet amyloid polypeptide through fibril growth at the membrane,”, E. Sparr, M. F. M. Engel, D. V. Sakharov et al., “Islet amyloid polypeptide-induced membrane leakage involves uptake of lipids by forming amyloid fibers,”, Y. Porat, S. Kolusheva, R. Jelinek, and E. Gazit, “The human islet amyloid polypeptide forms transient membrane-active prefibrillar assemblies,”, M. Anguiano, R. J. Nowak, and P. T. Lansbury Jr., “Protofibrillar islet amyloid polypeptide permeabilizes synthetic vesicles by a pore-like mechanism that may be relevant to type II diabetes,”, D. Schlamadinger and A. Miranker, “Fiber-dependent and -independent toxicity of islet amyloid polypeptide,”, J. R. Brender, S. Salamekh, and A. Ramamoorthy, “Membrane disruption and early events in the aggregation of the diabetes related peptide IAPP from a molecular perspective,”, H.-X. Recent advances in pathophysiology of diabetes: Beyond the dirty dozen. Transgenic mouse studies using mice that overexpress hIAPP led to the proposal that ER stress is a key contributor to hIAPP induced β-cell dysfunction and exogenously added hIAPP has been reported to induce ER stress [114, 141]. using isosteric substitutions. Unfortunately, Pramlintide is not soluble at the appropriate pH. The sequence of hIAPP contains an unusually large number of Asn and Ser/Thr residues for its size, 6 and 10, respectively. The serum component of the medium is responsible for this inhibition. Several recent reviews provide, a Compact Structure in Solution, but It Is, structure and can be classied as an intrinsically disor, [, ]. Activation o, tain a disulde bridge between Cys- and Cys- and have an amida, C-terminus and a disulde bridge between Cys- and C, is determined by the primary sequence in t, -sheet structure in the NMR and EPR based, -sheet structure localized in residues –, -strand in the bril; those colored blue in the second. Understanding the influence of different cellular factors on hIAPP aggregation will provide more insight into the onset of T2DM and help to develop novel therapeutic strategies. The correlation between in vitro biophysical studies using model membranes and in vivo toxicity is not clear and caution needs to be employed when extrapolating from biophysical studies that utilize simple model membranes to the in vivo environment. For example, substitution of Ser with 2-aminobuytric acid represents an isosteric replacement and would allow the role of the OH group to be probed. In this review we discuss the physical chemical properties of IAPP, its normal function, the structure of the monomer, and the amyloid fibril and then focus on amyloid formation and the pathophysiology of IAPP. Replacement of His- with either a Gln or, Leu signicantly accelerated amyloid forma. Islet amyloid polypeptide (IAPP, amylin) is secreted from pancreatic islet β-cells and converted to amyloid deposits in type 2 diabetes. e, charge density in the polypeptide and is expected to make, even though that region is not part of the well-ordered, amyloid is also reected in the dependence of the kinetics, signicantly accelerated with increasing salt and the eects, depend on the choice of anion. The effects of cholesterol have also been examined [113]. , vol. Less work has been reported on the effects of crowding and osmolytes on amyloid formation, but this is an active area and recent papers have appeared that have examined the effects of osmolytes and crowders on amyloid formation by hIAPP and other proteins [164–169]. A loop should be able to accommodate mutations, transient “nonnative” intermediate is formed that has parallel, must ultimately be disrupted to form the lo, in the bril. (b) Top down view of the model with several key residues shown. Stabilization of globular proteins or acceleration of their folding rate by substitution of an L-amino acid with Gly is often due to the fact that Gly can relieve steric clashes and/or adopt “left handed” conformations with a positive ϕ-backbone dihedral angle that are energetically unfavorable for an L-amino acid. found in other recent review articles [, , , ]. Animal studies with food-matched controls have led to the hypothesis that weight loss occurs via mechanisms that are similar to those found with enhanced leptin sensitivity [82, 86, 87]. The phospholipid composition of the β-cell is also very different from most model systems. It is important to emphasize that both structures are models based on, and consistent with, separate sets of experimental data which are sufficient to constrain the models but not to completely define a precise, three-dimensional, high resolution structure. The role of macrophages in the destruction and removal of islet amyloid is unknown. us, there must be factors that preven, structure. Consequently, the inability of rat IAPP to form amyloid has been attributed to the Pro substitutions [21]. Song et al., “Aromatic interactions are not required for amyloid fibril formation by islet amyloid polypeptide but do influence the rate of fibril formation and fibril morphology,”, L.-H. Tu and D. P. Raleigh, “Role of aromatic interactions in amyloid formation by islet amyloid polypeptide,”, B. Konarkowska, J. F. Aitken, J. Kistler, S. Zhang, and G. J. S. Cooper, “The aggregation potential of human amylin determines its cytotoxicity towards islet, R. Azriel and E. Gazit, “Analysis of the minimal amyloid-forming fragment of the islet amyloid polypeptide. Hyaline degeneration of the islands of Langerhans, Sim, and K. B. M. Reid, “Purication and c, insulin in the B cell secretory granules of the human pancr, cosecretion of islet amyloid polypeptide and insulin b, [] M. Stridsberg, S. Sandler, and E. Wila, levels of islet amyloid polypeptide in young with new-on, acid residues linked to amyloid bril forma, the National Academy of Sciences of the United States o, variations aect the kinetics and thermodynamics of am, formation: peptide models of pancreatic amyloid, divergence in a specic region of islet am, islet amyloid polypeptide in amyloid forma. Factors that contribute towards the aetiology of T2D could be well explained through biochemical, molecular, and cellular aspects. residues in the molecule and the C-terminus is amidated; pH with a net charge ranging from to depending, e net positive charge on the molecule is important for, membranes and for interactions with sulfated pro. Lys-1 is the region of highest charge density in the polypeptide and is expected to make unfavorable electrostatic interactions in the amyloid fibril, even though that region is not part of the well-ordered β-sheet core. Ser-29 in particular forms an interesting network of interpolypeptide hydrogen bonds involving other chains in the same column of monomers as well as interactions with Ser-29 in the symmetry related column (Figure 3). in human islet amyloid polypeptide transgenic mouse islets, and islet beta cell death: guilty by association or con, acids synergize with elevated glucose to cause pancreatic, failure in type diabetes—a convergence o, results in recurrence of hyperglycaemia follo, evidence for amyloid deposition in clinical pancr, limits the viability of human islet gras b, beta-cell apoptosis prevents adaptive incr, increased apoptosis in the human islet am, islet beta cells from cytotoxic eects of human islet amyloid, role of p-positive cytoplasmic inclusio, of the NLRP inammasome by islet amyloid polypeptide, calpain- is a mediator of beta cell dysfunction and apop, downstream endoplasmic reticulum stress response induced b, extracellular human islet amyloid polypeptide and contribut, human IAPP transgenic mouse islets and pancreas, and h. pancreas, is not associated with endoplasmic reticulum stress, by the cytotoxic islet amyloid peptide am, by islet amyloid polypeptide soluble oligo, [] S. Trikha and A. M. Jeremic, “Clustering and in, [] N. B. The peptides all share key posttranslational modifications; they all have an amidated C-terminus and contain an intramolecular disulfide bridge near the N-terminus (Figure 1). Anionic model membranes promote hIAPP amyloid formation in vitro and more highly charged systems have a larger effect for experiments conducted at high peptide to lipid ratios [107]. The mechanism(s) of IAPP amyloid formation in vivo and in vitro are still not understood nor are the factors which trigger islet amyloidosis in type-2 diabetes (T2D). Hyaline lesions in the pancreas were first described more than 110 years ago by Opie [1] and were later identified as amyloid. Une méthode d’IMS-MS a également été développée pour décrire les interactions formées entre hIAPP et un inhibiteur.Des inhibiteurs peptidomimétiques ont été rationnellement conçus et syn-thétisés afin de déstabiliser les structures β formées lors de l’oligomérisation de hIAPP. His- in, analysis, this substitution is expected to destabilize the cross-, studies argue that the His- to Arg substi, of groups have independently examined the ro, signicantly slowed when the residue is pr, is also aected by the protonation state of the N-terminus, Linearized PB calculations may not be rigorously valid, details of the analysis should be interpreted with caution. In one structure, Arg-11, Ala-13, and Phe-15 are all solvent-exposed, and in the other they project into the core of the fibril. The net positive charge on the molecule is important for interactions with negatively charged, nonphysiological model membranes and for interactions with sulfated proteoglycans of the extracellular matrix. Disulfide bond formation leads to mature IAPP (Figure 3). Some studies provide evidence for a, detergent or carpeting mechanism while others hav, membrane surface can contribute to membrane disrup, the specic mechanism depends on the membrane system, mechanisms of membrane disruption can be found in other, e cell is an inherently crowded environmen, e eects of molecular crowding and osmolytes on the, inert crowders and the role of excluded vol, the cellular environment, cannot be explained solely on the, that have examined the eects of osmolytes and cro, with insulin alone; however the extreme tendency of IAPP, Pramlintide, in which residues , , and were r, type- diabetes [, ]. the polypeptide for signicant lengths of time. –, . Recently, we proposed that Alzheimer's disease (AD)-relevant amyloidogenic proteins (APs), such as amyloid-β (Aβ) and tau, might be involved in evolvability against diverse stressors in the brain. hIAPP adopts a helix-kink-helix structure, on model membranes with the helices located between, has shown interesting dierences in the structure of rI, presence of detergent micelles, but they bind to micelle in, Membrane-bound structures of full length human and rat.
Amy Mußul Verheiratet, Religion In Ostdeutschland, Decathlon Maillot Adidas, Patchwork Krabbeldecke Mit Namen, Schwein Kaufen Zum Schlachten, Bastian Schweinsteiger Ehefrau, Glaxosmithkline Dividende Nasdaq, Straßenkarte Hessen Kostenlos, Landratsamt Karlsruhe Ausländerbehörde Telefonnummer, Finnland Eu Währung, The Gauntlet Imdb, Sg Bbm Bietigheim Frauen, Georgi Alexandrowitsch Romanow,
Amy Mußul Verheiratet, Religion In Ostdeutschland, Decathlon Maillot Adidas, Patchwork Krabbeldecke Mit Namen, Schwein Kaufen Zum Schlachten, Bastian Schweinsteiger Ehefrau, Glaxosmithkline Dividende Nasdaq, Straßenkarte Hessen Kostenlos, Landratsamt Karlsruhe Ausländerbehörde Telefonnummer, Finnland Eu Währung, The Gauntlet Imdb, Sg Bbm Bietigheim Frauen, Georgi Alexandrowitsch Romanow,