Amyloid


Amyloids are aggregates of proteins characterised by a fibrillar morphology of typically 7–13 nm in diameter, a β-sheet secondary structure and ability to be stained by particular dyes, such as Congo red. In the human body, amyloids have been linked to the development of various diseases. Pathogenic amyloids form when previously healthy proteins lose their normal structure and physiological functions and form fibrous deposits within and around cells. These protein misfolding and deposition processes disrupt the healthy function of tissues and organs.
Such amyloids have been associated with more than 50 human diseases, including amyloidosis, and may play a role in some neurodegenerative diseases. Some of these diseases are mainly sporadic and only a few cases are familial. Others are only familial. Some result from medical treatment. Prions are an infectious form of amyloids that can act as a template to convert other non-infectious forms. Amyloids may also have normal biological functions; for example, in the formation of fimbriae in some genera of bacteria, transmission of epigenetic traits in fungi, as well as pigment deposition and hormone release in humans.
Amyloids have been known to arise from many different proteins. These polypeptide chains generally form β-sheet structures that aggregate into long fibers; however, identical polypeptides can fold into multiple distinct amyloid conformations. The diversity of the conformations may have led to different forms of the prion diseases.
An unusual secondary structure named α sheet has been proposed as the toxic constituent of amyloid precursor proteins, but this idea is not widely accepted at present.

Definition

The name amyloid comes from the early mistaken identification by Rudolf Virchow of the substance as starch, based on crude iodine-staining techniques. For a period, the scientific community debated whether or not amyloid deposits are fatty deposits or carbohydrate deposits until it was finally found that they are, in fact, deposits of albumin-like proteinaceous material.
  • The classical, histopathological definition of amyloid is an extracellular, proteinaceous fibrillar deposit exhibiting β-sheet secondary structure and identified by apple-green birefringence when stained with congo red under polarized light. These deposits often recruit various sugars and other components such as serum amyloid P component, resulting in complex, and sometimes inhomogeneous structures. Recently this definition has come into question as some classic, amyloid species have been observed in distinctly intracellular locations.
  • A more recent, biophysical definition is broader, including any polypeptide that polymerizes to form a cross-β structure, in vivo or in vitro, inside or outside cells. Microbiologists, biochemists, biophysicists, chemists and physicists have largely adopted this definition, leading to some conflict in the biological community over an issue of language.

    Proteins forming amyloids in diseases

To date, 37 human proteins have been found to form amyloid in pathology and be associated with well-defined diseases. The International Society of Amyloidosis classifies amyloid fibrils and their associated diseases based upon associated proteins. A table is included below.
ProteinDiseasesOfficial abbreviation
β amyloid peptide from Amyloid precursor proteinAlzheimer's disease, Hereditary cerebral haemorrhage with amyloidosis
α-synucleinParkinson's disease, Parkinson's disease dementia, Dementia with Lewy bodies, Multiple system atrophyAαSyn
PrPScTransmissible spongiform encephalopathy APrP
Microtubule-associated protein tauVarious forms of tauopathies ATau
Huntingtin exon 1Huntington's diseaseHTTex1
ABri peptideFamilial British dementiaABri
ADan peptideFamilial Danish dementiaADan
Fragments of immunoglobulin light chainsLight chain amyloidosisAL
Fragments of immunoglobulin heavy chainsHeavy chain amyloidosisAH
full length of N-terminal fragments of Serum amyloid A proteinAA amyloidosisAA
TransthyretinSenile systemic amyloidosis, Familial amyloid polyneuropathy, Familial amyloid cardiomyopathy, Leptomeningeal amyloidosisATTR
β-2 microglobulinDialysis related amyloidosis, Hereditary visceral amyloidosis Aβ2M
N-terminal fragments of Apolipoprotein AIApoAI amyloidosisAApoAI
C-terminally extended Apolipoprotein AIIApoAII amyloidosisAApoAII
N-terminal fragments of Apolipoprotein AIVApoAIV amyloidosisAApoAIV
Apolipoprotein C-IIApoCII amyloidosisAApoCII
Apolipoprotein C-IIIApoCIII amyloidosisAApoCIII
fragments of GelsolinFamilial amyloidosis, Finnish typeAGel
LysozymeHereditary non-neuropathic systemic amyloidosisALys
fragments of Fibrinogen α chainFibrinogen amyloidosisAFib
N-terminally truncated Cystatin CHereditary cerebral hemorrhage with amyloidosis, Icelandic typeACys
IAPP Diabetes mellitus type 2, InsulinomaAIAPP
CalcitoninMedullary carcinoma of the thyroidACal
Atrial natriuretic factorCardiac arrhythmias, Isolated atrial amyloidosisAANF
ProlactinPituitary prolactinomaAPro
InsulinInjection-localized amyloidosisAIns
Lactadherin / MedinAortic medial amyloidosisAMed
Lactotransferrin / LactoferrinGelatinous drop-like corneal dystrophyALac
Odontogenic ameloblast-associated proteinCalcifying epithelial odontogenic tumorsAOAAP
Pulmonary surfactant-associated protein C Pulmonary alveolar proteinosisASPC
Leukocyte cell-derived chemotaxin-2 Renal LECT2 amyloidosisALECT2
Galectin-7Lichen amyloidosis, Macular amyloidosisAGal7
CorneodesmosinHypotrichosis simplex of the scalpACor
C-terminal fragments of TGFBI/KeratoepithelinLattice corneal dystrophy type I, Lattice corneal dystrophy type 3A, Lattice corneal dystrophy Avellino typeAKer
Semenogelin-1 Seminal vesicle amyloidosisASem1
Proteins S100A8/A9Prostate cancernone
EnfuvirtideInjection-localized amyloidosisAEnf

Non-disease and functional amyloids

Many examples of non-pathological amyloid with a well-defined physiological role have been identified in various organisms, including human. These may be termed as functional or physiological or native amyloid.
  • Functional amyloid in Homo sapiens:
  • * Intralumenal domain of melanocyte protein PMEL
  • * Peptide/protein hormones stored as amyloids within endocrine secretory granules
  • * Receptor-interacting serine/threonine-protein kinase 1/3
  • * Fragments of prostatic acid phosphatase and semenogelins
  • Functional amyloid in other organisms:
  • * Curli fibrils produced by E. coli, ''Salmonella, and a few other members of the Enterobacteriales. The genetic elements encoding the curli system are phylogenetic widespread and can be found in at least four bacterial phyla. This suggest that many more bacteria may express curli fibrils.
  • * GvpA, forming the walls of particular Gas vesicles, i.e. the buoyancy organelles of aquatic archaea and eubacteria
  • * Fap fibrils in various species of Pseudomonas
  • * Chaplins from Streptomyces coelicolor
  • * Spidroin from Trichonephila edulis
  • * Hydrophobins from Neurospora crassa and other fungi
  • * Fungal cell adhesion proteins forming cell surface amyloid regions with greatly increased binding strength
  • * Environmental biofilms according to staining with amyloid specific dyes and antibodies.
  • * Tubular sheaths encasing Methanosaeta thermophila filaments
  • Functional amyloid acting as prions
  • * Several yeast prions are based on an infectious amyloid, e.g. ; ; or ; and
  • * Prion HET-s from Podospora anserina
  • * Neuron-specific isoform of CPEB from Aplysia californica''

    Structure

Amyloids are formed of long unbranched fibers that are characterized by an extended β-sheet secondary structure in which individual β strands are arranged in an orientation perpendicular to the long axis of the fiber. Such a structure is known as cross-β structure. Each individual fiber may be 7–13 nanometres in width and a few micrometres in length. The main hallmarks recognised by different disciplines to classify protein aggregates as amyloid is the presence of a fibrillar morphology with the expected diameter, detected using transmission electron microscopy or atomic force microscopy, the presence of a cross-β secondary structure, determined with circular dichroism, FTIR, solid-state nuclear magnetic resonance, X-ray crystallography, or X-ray fiber diffraction, and an ability to stain with specific dyes, such as Congo red, thioflavin T or thioflavin S.
The term "cross-β" was based on the observation of two sets of diffraction lines, one longitudinal and one transverse, that form a characteristic "cross" pattern. There are two characteristic scattering diffraction signals produced at 4.7 and 10 Å, corresponding to the interstrand and stacking distances in β sheets. The "stacks" of β sheet are short and traverse the breadth of the amyloid fibril; the length of the amyloid fibril is built by aligned β-strands. The cross-β pattern is considered a diagnostic hallmark of amyloid structure.
Amyloid fibrils are generally composed of 1–8 protofilaments, each 2–7 nm in diameter, that interact laterally as flat ribbons that maintain the height of 2–7 nm and are up to 30 nm wide; more often protofilaments twist around each other to form the typically 7–13 nm wide fibrils. Each protofilament possesses the typical cross-β structure and may be formed by 1–6 β-sheets stacked on each other. Each individual protein molecule can contribute one to several β-strands in each protofilament and the strands can be arranged in antiparallel β-sheets, but more often in parallel β-sheets. Only a fraction of the polypeptide chain is in a β-strand conformation in the fibrils, the remainder forms structured or unstructured loops or tails.
For a long time our knowledge of the atomic-level structure of amyloid fibrils was limited by the fact that they are unsuitable for the most traditional methods for studying protein structures. Recent years have seen progress in experimental methods, including solid-state NMR spectroscopy and cryo-electron microscopy. Combined, these methods have provided 3D atomic structures of amyloid fibrils formed by amyloid β peptides, α-synuclein, tau, and the FUS protein, associated with various neurodegenerative diseases.
X-ray diffraction studies of microcrystals revealed atomistic details of core region of amyloid, although only for simplified peptides having a length remarkably shorter than that of peptides or proteins involved in disease. The crystallographic structures show that short stretches from amyloid-prone regions of amyloidogenic proteins run perpendicular to the filament axis, consistent with the "cross-β" feature of amyloid structure. They also reveal a number of characteristics of amyloid structures – neighboring β-sheets are tightly packed together via an interface devoid of water, with the opposing β-strands slightly offset from each other such that their side-chains interdigitate. This compact dehydrated interface created was termed a steric-zipper interface. There are eight theoretical classes of steric-zipper interfaces, dictated by the directionality of the β-sheets and symmetry between adjacent β-sheets. A limitation of X-ray crystallography for solving amyloid structure is represented by the need to form microcrystals, which can be achieved only with peptides shorter than those associated with disease.
Although bona fide amyloid structures always are based on intermolecular β-sheets, different types of "higher order" tertiary folds have been observed or proposed. The β-sheets may form a β-sandwich, or a β-solenoid which may be either β-helix or β-roll. Native-like amyloid fibrils in which native β-sheet containing proteins maintain their native-like structure in the fibrils have also been proposed. There are few developed ideas on how the complex backbone topologies of disulfide-constrained proteins, which are prone to form amyloid fibrils, adopt the amyloid β-sheet motif. The presence of multiple constraints significantly reduces the accessible conformational space, making computational simulations of amyloid structures more feasible.
One complicating factor in studies of amyloidogenic polypeptides is that identical polypeptides can fold into multiple distinct amyloid conformations. This phenomenon is typically described as amyloid polymorphism.