Molecular properties and fibril ultrastructure of types II and XI collagens in cartilage of mice expressing exclusively the 伪1(IIA) collagen isoform

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• 作者：Audrey McAlinden^a ; ^b ; Geoffrey Traeger^c ; Uwe Hansen^d ; Mary Ann Weis^c ; Soumya Ravindran^a ; Louisa Wirthlin^a ; David R. Eyre^c ; Russell J. Fernandes^c ; ^{rjf@u.washington.edu" class="auth_mail}
• 关键词：Type IIA procollagen ; Alternative splicing ; Cartilage ; Collagen cross-links ; Heteropolymer ; Collagen fibrils ; Extracellular matrix ; Post-translational modifications
• 刊名：Matrix Biology
• 出版年：February 2014
• 年：2014
• 卷：34
• 期：Complete
• 页码：105-113
• 全文大小：2051 K

文摘

Until now, no biological tools have been available to determine if a cross-linked collagen fibrillar network derived entirely from type IIA procollagen isoforms, can form in the extracellular matrix (ECM) of cartilage. Recently, homozygous knock-in transgenic mice (Col2a1^+ ex2, ki/ki) were generated that exclusively express the IIA procollagen isoform during post-natal development while type IIB procollagen, normally present in the ECM of wild type mice, is absent. The difference between these Col2a1 isoforms is the inclusion (IIA) or exclusion (IIB) of exon 2 that is alternatively spliced in a developmentally regulated manner. Specifically, chondroprogenitor cells synthesize predominantly IIA mRNA isoforms while differentiated chondrocytes produce mainly IIB mRNA isoforms. Recent characterization of the Col2a1^+ ex2 mice has surprisingly shown that disruption of alternative splicing does not affect overt cartilage formation. In the present study, biochemical analyses showed that type IIA collagen extracted from ki/ki mouse rib cartilage can form homopolymers that are stabilized predominantly by hydroxylysyl pyridinoline (HP) cross-links at levels that differed from wild type rib cartilage. The findings indicate that mature type II collagen derived exclusively from type IIA procollagen molecules can form hetero-fibrils with type XI collagen and contribute to cartilage structure and function. Heteropolymers with type XI collagen also formed. Electron microscopy revealed mainly thin type IIA collagen fibrils in ki/ki mouse rib cartilage. Immunoprecipitation and mass spectrometry of purified type XI collagen revealed a heterotrimeric molecular composition of 伪1(XI)伪2(XI)伪1(IIA) chains where the 伪1(IIA) chain is the IIA form of the 伪3(XI) chain. Since the N-propeptide of type XI collagen regulates type II collagen fibril diameter in cartilage, the retention of the exon 2-encoded IIA globular domain would structurally alter the N-propeptide of type XI collagen. This structural change may subsequently affect the regulatory function of type XI collagen resulting in the collagen fibril and cross-linking differences observed in this study.