Isolation and Characterisation of Major and Minor Collagens from Hyaline Cartilage of Hoki (Macruronus novaezelandiae).

Cumming Cumming,Hofman Hofman,Hall Hall

doi:10.3390/md17040223

Abstract

The composition and properties of collagen in teleost (bony fish) cartilage have never been studied. In this study, we aimed to identify and characterise all collagen species in the nasal cartilage of hoki (Macruronus novaezelandiae). Four native collagen species were extracted using two techniques, and isolated with differential salt precipitation. We were able to assign the identity of three of these collagen species on the basis of solubility, SDS-PAGE and amino acid analyses. We found that hoki cartilage contains the major collagen, type II, and the minor collagens, type IX and type XI, which are homologous to those found in mammal and chicken cartilage. Using these extraction protocols, we also isolated a full-length type IX collagen from cartilage for the first time. In addition, we detected a 90 kDa, highly glycosylated collagen that has not been identified in any other species. For each isolate, structural and biochemical characterisations were performed using circular dichroism and Fourier transform infrared spectroscopy analyses, and the thermal denaturation properties were determined. Our results showed that the properties of hoki cartilage-derived collagens are similar to those of collagens in mammalian cartilage, indicating that teleost cartilage could provide biological ingredients for the development of biomaterials to treat cartilage-related illnesses.

Highlights

Collagens and proteoglycans are the main proteins in the extra cellular matrix of cartilage and, together with water, they give cartilage its strength and elastic properties [1]
Proteoglycans were removed using guanidinium hydrochloride (GuHCl), which was necessary for effective protein isolation from cartilage
Identification and characterisation of the major and minor collagens that make up fibrils in terrestrial animals are well established [61]

Summary

Introduction

Collagens and proteoglycans are the main proteins in the extra cellular matrix of cartilage and, together with water, they give cartilage its strength and elastic properties [1]. Type II collagen is a fibril-forming collagen found exclusively in cartilage, vitreous humour, and the notochord. It is a homotrimeric molecule consisting of three identical alpha chains ([α1(II)]3 ), each approximately. 120 kDa. Type XI collagen is a heterotrimeric fibril-forming collagen with three distinct alpha chains: α1(XI), α2(XI) and α3(XI)] [3]. The α3(XI) chain has the same primary sequence as α1(II) but differs in its degree of glycosylation. Little is known about type XI collagen. It is believed to play a key role in initiating fibrillogenesis and its presence in the fibril has been shown to restrict fibril width [4]

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