Golgi reassembly stacking protein 55 interacts with membrane‐type (MT) 1‐matrix metalloprotease (MMP) and furin and plays a role in the activation of the MT1‐MMP zymogen
Authors: Roghi Christian , Jones Louise , Gratian Matthew , English William R. , Murphy Gillian 
Published: July 2010

Membrane‐type 1 matrix metalloproteinase (MT1‐MMP) is a proteinase involved in the remodelling of extracellular matrix and the cleavage of a number of substrates. MT1‐MMP is synthesized as a zymogen that requires intracellular post‐translational cleavage to gain biological activity. Furin, a member of the pro‐protein convertase family, has been implicated in the proteolytic removal of the MT1‐MMP prodomain sequence. In the present study, we demonstrate a role for the peripheral Golgi matrix protein GRASP55 in the furin‐dependent activation of MT1‐MMP. MT1‐MMP and furin were found to co‐localize with Golgi reassembly stacking protein 55 (GRASP55). Further analysis revealed that GRASP55 associated with the cytoplasmic domain of both proteases and that the LLY573 motif in the MT1‐MMP intracellular domain was crucial for the interaction with GRASP55. Overexpression of GRASP55 was found to enhance the formation of a complex between MT1‐MMP and furin. Finally, we report that disruption of the interaction between GRASP55 and furin led to a reduction in pro‐MT1‐MMP activation. Taken together, these data suggest that GRASP55 may function as an adaptor protein coupling MT1‐MMP with furin, thus leading to the activation of the zymogen.

Structured digital abstract

 MINT‐7897990: Furin (uniprotkb:P09958) and GRASP55 (uniprotkb:Q9H8Y8) colocalize (MI:0403) by fluorescence microscopy (MI:0416)

 MINT‐7897801: GRASP55 (uniprotkb:Q9R064) physically interacts (MI:0915) with MT2‐MMP (uniprotkb:P51511) by two hybrid (MI:0018)

 MINT‐7897821: GRASP55 (uniprotkb:Q9R064) physically interacts (MI:0915) with MT3‐MMP (uniprotkb:P51512) by two hybrid (MI:0018)

 MINT‐7897577: GRASP55 (uniprotkb:Q9R064) and MT1‐MMP (uniprotkb:P50281) colocalize (MI:0403) by fluorescence microscopy (MI:0416)

 MINT‐7897366: MT1‐MMP (uniprotkb:P50281) physically interacts (MI:0915) with GRASP55 (uniprotkb:Q9H8Y8) by anti bait coimmunoprecipitation (MI:0006)

 MINT‐7897617, MINT‐7897659, MINT‐7897681, MINT‐7897702, MINT‐7897725, MINT‐7898032, MINT‐7898011, MINT‐7897907, MINT‐7897884: GRASP55 (uniprotkb:Q9R064) physically interacts (MI:0915) with MT1‐MMP (uniprotkb:P50281) by two hybrid (MI:0018)

 MINT‐7898002: MT1‐MMP (uniprotkb:P50281) physically interacts (MI:0914) with Furin (uniprotkb:P09958) by anti bait coimmunoprecipitation (MI:0006)

 MINT‐7897500: MT1‐MMP (uniprotkb:P50281) and Giantin (uniprotkb:Q14789) colocalize (MI:0403) by fluorescence microscopy (MI:0416)

 MINT‐7897750, MINT‐7897394: GRASP55 (uniprotkb:Q9R064) physically interacts (MI:0915) with MT1‐MMP (uniprotkb:P50281) by anti tag coimmunoprecipitation (MI:0007)

 MINT‐7897562: MT1‐MMP (uniprotkb:P50281) and GRASP55 (uniprotkb:Q9H8Y8) colocalize (MI:0403) by fluorescence microscopy (MI:0416)

 MINT‐7897512: TGN46 (uniprotkb:O43493) and MT1‐MMP (uniprotkb:P50281) colocalize (MI:0403) by fluorescence microscopy (MI:0416)

 MINT‐7897921, MINT‐7897975: GRASP55 (uniprotkb:Q9R064) physically interacts (MI:0915) with Furin (uniprotkb:P09958) by two hybrid (MI:0018)

 MINT‐7898052, MINT‐7897410: MT1‐MMP (uniprotkb:P50281) physically interacts (MI:0915) with GRASP55 (uniprotkb:Q9R064) by anti bait coimmunoprecipitation (MI:0006)

 MINT‐7897951: GRASP55 (uniprotkb:Q9R064) physically interacts (MI:0915) with PC7 (uniprotkb:Q16549) by two hybrid (MI:0018)

 MINT‐7897866: GRASP55 (uniprotkb:Q9R064) physically interacts (MI:0915) with MT5‐MMP (uniprotkb:Q9Y5R2) by two hybrid (MI:0018)

 MINT‐7897633: GRASP55 (uniprotkb:Q9R064) physically interacts (MI:0915) with TGFA (uniprotkb:P01135) by two hybrid (MI:0018)

 MINT‐7897551: GRASP55 (uniprotkb:Q9H8Y8) and Giantin (uniprotkb:Q14789) colocalize (MI:0403) by fluorescence microscopy (MI:0416)

 MINT‐7897938: GRASP55 (uniprotkb:Q9R064) physically interacts (MI:0915) with PC5/6B (uniprotkb:Q04592) by two hybrid (MI:0018)