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Glycosylation modulates melanoma cell ?2?1 and ?3?1 integrin interactions with type IV collagen.

ABSTRACT: Although type IV collagen is heavily glycosylated, the influence of this post-translational modification on integrin binding has not been investigated. In the present study, galactosylated and nongalactosylated triple-helical peptides have been constructed containing the ?1(IV)382-393 and ?1(IV)531-543 sequences, which are binding sites for the ?2?1 and ?3?1 integrins, respectively. All peptides had triple-helical stabilities of 37 °C or greater. The galactosylation of Hyl(393) in ?1(IV)382-393 and Hyl(540) and Hyl(543) in ?1(IV)531-543 had a dose-dependent influence on melanoma cell adhesion that was much more pronounced in the case of ?3?1 integrin binding. Molecular modeling indicated that galactosylation occurred on the periphery of ?2?1 integrin interaction with ?1(IV)382-393 but right in the middle of ?3?1 integrin interaction with ?1(IV)531-543. The possibility of extracellular deglycosylation of type IV collagen was investigated, but no ?-galactosidase-like activity capable of collagen modification was found. Thus, glycosylation of collagen can modulate integrin binding, and levels of glycosylation could be altered by reduction in expression of glycosylation enzymes but most likely not by extracellular deglycosylation activity.

SUBMITTER: Stawikowski MJ

PROVIDER: S-EPMC4118119 | biostudies-literature | 2014 Aug

REPOSITORIES: biostudies-literature

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Glycosylation modulates melanoma cell α2β1 and α3β1 integrin interactions with type IV collagen.

Stawikowski Maciej J MJ Aukszi Beatrix B Stawikowska Roma R Cudic Mare M Fields Gregg B GB

The Journal of biological chemistry 20140623 31

Although type IV collagen is heavily glycosylated, the influence of this post-translational modification on integrin binding has not been investigated. In the present study, galactosylated and nongalactosylated triple-helical peptides have been constructed containing the α1(IV)382-393 and α1(IV)531-543 sequences, which are binding sites for the α2β1 and α3β1 integrins, respectively. All peptides had triple-helical stabilities of 37 °C or greater. The galactosylation of Hyl(393) in α1(IV)382-393 ...[more]

PMID: 24958723

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Project description:Type X collagen is a short-chain non-fibrillar collagen that is deposited exclusively at sites of new bone formation. Although this collagen has been implicated in chondrocyte hypertrophy and endochondral ossification, its precise function remains unclear. One possible function could be to regulate the processes of chondrocyte hypertrophy through direct cell-type X collagen interactions. Adhesions of embryonic chick chondrocytes, and cell lines with known expression of collagen-binding integrins (MG63 and HOS), were assayed on chick type X collagen substrates, including the native, heat-denatured and pepsin-digested collagen, and the isolated C-terminal non-collagenous (NC1) domain. Type X collagen supported the greatest level of adhesion for all cell types tested. The involvement of the alpha2beta1 integrin in type X collagen-cell interaction was demonstrated by adhesion studies in the presence of Mg(2+) and Ca(2+) ions and integrin-function-blocking antibodies. Cells expressing alpha2beta1 integrin (chick chondrocytes and MG63 cells) also adhered to heat-denatured type X collagen and the isolated NC1 domain; however, removal of the non-collagenous domains by limited pepsinization of type X collagen resulted in very low levels of adhesion. Both focal contacts and actin stress-fibre formation were apparent in cells plated on type X collagen. The presence of alpha2 and beta1 integrin subunits in isolated chondrocytes and epiphyseal cartilage was also confirmed by immunolocalization. Our results demonstrate, for the first time, that type X collagen is capable of interacting directly with chondrocytes and other cells, primarily via alpha2beta1 integrin. These findings are atypical from the fibrillar collagen-cell interactions via collagen binding integrins in that: (1) the triple-helical conformation is not strictly required for cell adhesion; (2) the NC1 domain is also involved in the adhesion of alpha2beta1-expressing cells. These data form the basis for further studies into the mechanism and biological significance of type X collagen deposition in the growth plate.

Dataset Information

Glycosylation modulates melanoma cell ?2?1 and ?3?1 integrin interactions with type IV collagen.

Publications

Glycosylation modulates melanoma cell α2β1 and α3β1 integrin interactions with type IV collagen.

Similar Datasets

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