Amyloid from a histochemical perspective. A review of the structure, properties and types of amyloid, and a proposed staining mechanism for Congo red staining. (17th November 2018)

• Record Type: Journal Article
• Authors: Dapson, Richard W.
• Is Part Of: Biotechnic & histochemistry. Volume 93:Number 8(2018)
• Page Start: 543
• Page End: 556
• Publication Date: 2018-11-17
• Subjects: Amyloid -- atomic structure -- ß-sheet -- ß-strand -- bonding mechanisms -- bonding sites -- Congo red -- Debye forces -- histochemistry -- hydrogen bonding -- ionic bonding -- Keesom forces -- London forces -- van der Waals forces; Histochemistry -- Periodicals; Stains and staining (Microscopy) -- Periodicals; Biotechnology -- Periodicals; 572
• DOI: 10.1080/10520295.2018.1528385
• Languages: English
• ISSNs: 1052-0295

Abstract

ABSTRACT: Amyloid is a diverse group of unrelated peptides or proteins that have positive functionality or are associated with various pathologies. Despite vast differences, all amyloids share several features that together uniquely define the group. 1) All amyloids possess a characteristic cross- ß pattern with X-ray diffraction typical of ß -sheet secondary protein structures. 2) All amyloids are birefringent and dichroic under polarizing microscopy after staining with Congo red, which indicates a crystalline-like (ordered) structure. 3) All amyloids cause a spectral shift in the peak wavelength of Congo red with conventional light microscopy due to perturbation of π electrons of the dye. 4) All amyloids show heightened intensity of fluorescence with Congo red, which suggests an unusual degree of packing of the dye onto the substrate. The ß portion of amyloid molecules, the only logical substrate for specific Congo red staining under histochemical conditions, consists of a stack of ß -sheets laminated by hydrophilic and hydrophobic interactions between adjacent pairs. Only the first and last ß -sheets are accessible to dyes. Each sheet is composed of numerous identical peptides running across the width of the sheet and arranged in parallel with side chains in register over the length of the fibril. Two sets of grooves are bordered by side chains. X grooves run perpendicular to the long axis of the fibril; these grooves are short (the width of the sheet) and number in the hundreds or thousands. Y grooves are parallel with the long axis. Each groove runs the entire length of the fibril, but there are very few of them. While Congo red is capable of ionic bonding with proteins via two sulfonic acid groups, physical constraints on the staining solution preclude ionic interactions. Hydrogen bonding between dye amine groups and peptide carbonyls is the most likely primary bonding mechanism, because all ß -sheets possess backbone carbonyls. Various amino acid residues may form secondary bonds to the dye via any of three van der Waals forces. It is possible that Congo red binds within the Y grooves, but that would not produce the characteristic staining features that are the diagnostic hallmarks of amyloid. Binding in the X grooves would produce a tightly packed series of dye molecules over the entire length of the fibril. This would account for the signature staining of amyloid by Congo red: dichroic birefringence, enhanced intensity of fluorescence and a shift in visible absorption wavelength.