Membrane‐type 1 MMP (MMP‐14) cleaves at three sites in the aggrecan interglobular domain

FEBS Letters

Last, Karena⁴  Fujii, Yutaka³  Okada, Yasunori¹  Seiki, Motoharu²  Fosang, Amanda J⁴ 

[1] Department of Pathology, School of Medicine, Keio University, Shinjuku-ku, Tokyo 160-0016, Japan;Department of Cancer Research, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-0071, Japan;Department of Chemistry, Fukui Medical University, Fukui 910-1193, Japan;Orthopaedic Molecular Biology Research Unit, Melbourne University, Department of Paediatrics, Royal Children's Hospital, Parkville, 3052, Australia

关键词: Matrix metalloproteinase;    Membrane-type matrix metalloproteinase;    Aggrecan;    Neo-epitope;    Arthritis;    IGD;    interglobular domain of aggrecan;    MMPs;    matrix metalloproteinases;    MT-MMPs;    membrane-type matrix metalloproteinases;    OA;    osteoarthritis;    IL-1;    interleukin-1;    TNF;    tumour necrosis factor;    HRP;    horseradish peroxidase;    PVDF;    polyvinylidene difluoride;    AEBSF;    [4-(2-aminoethyl)benzene]sulfonylfluoride;    PBS;    phosphate-buffered saline;    ΔMT1;    deletion mutant of MT1-MMP lacks the COOH-terminal transmembrane and cytoplasmic domain (Δ Ala536–Val582) (see [20]);   

DOI  :  10.1016/S0014-5793(98)00667-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

An aggrecan G1-G2 substrate was used to determine sites within the interglobular domain that were susceptible to cleavage by MT1-MMP. Degradation products were identified by Western blotting with neo-epitope antibodies specific for MMP-derived N- and C-terminal sequences. The results showed that MT1-MMP cleaved at the N₃₄₁-F₃₄₂ and D₄₄₁-L₄₄₂ bonds, as shown for other MMPs, and also at a site 13 amino acids C-terminal to the N₃₄₁-F₃₄₂ site. The G2 product of this additional cleavage was identified by sequence analysis and revealed an N-terminus commencing T₃₅₅VxxPDVELPLP. The data are consistent with MT1-MMP cleavage at three sites in the aggrecan interglobular domain; one at N₃₄₂-F₃₄₂, a second at D₄₄₁-L₄₄₂ and a third at Q₃₅₄-T₃₅₅.

【 授权许可】

Unknown