Domain 5 of high molecular weight kininogen inhibits collagen-mediated cancer cell adhesion and invasion in association with ¦Á-actinin-4

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• 作者：Tsunetoshi Hatoh^a ; ^b ; ^c ; Toshinaga Maeda^a ; Keisuke Takeuchi^a ; Osamu Ogikubo^b ; ^d ; Susumu Uchiyama^e ; Takanobu Otsuka^b ; Iwao Ohkubo^a ; ^f ; ^{ohkubo@tenshi.ac.jp} ; Hisakazu Ogita^a ; ^{hogita@belle.shiga-med.ac.jp}
• 关键词：HK ; high molecular weight kininogen ; D5H ; domain 5 of high molecular weight kininogen ; MEME ; Minimum essential medium Eagle ; GST ; glutathione-S-transferase ; Col-I ; collagen type-I ; PBS ; phosphate-buffered saline ; BSA ; bovine serum albumin ; PMSF ; phenylmet
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2012
• 出版时间：26 October, 2012
• 年：2012
• 卷：427
• 期：3
• 页码：497-502
• 全文大小：386 K

文摘

High molecular weight kininogen (HK) is a plasma glycoprotein with multiple functions, including the regulation of coagulation. We previously demonstrated that domain 5 (D5_H), a functional domain of HK, and its derived peptides played an important role in the vitronectin-mediated suppression of cancer cell adhesion and invasion. However, the underlying mechanisms of the D5_H-mediated suppressive effects remain to be elucidated. Here, we showed that D5_H and its derivatives inhibited the collagen-mediated cell adhesion and invasion of human osteosarcoma MG63 cells. Using purified D5_H fused to glutathione-S-transferase (GST) and D5_H-derived peptides for column chromatography, an actin-binding protein, ¦Á-actinin-4, was identified as a binding protein of D5_H with high-affinity for P-5m, a core octapeptide of D5_H. Immunofluorescence microscopy demonstrated that D5_H co-localized with ¦Á-actinin-4 inside MG63 cells. In addition, exogenous GST-D5_H added to the culture media was transported into MG63 cells, although GST alone as a control was not. As ¦Á-actinin-4 regulates actin polymerization necessary for cell adhesion and is related to the integrin-dependent attachment of cells to the extracellular matrix, our results suggest that D5_H may modulate cell adhesion and invasion together with actinin-4.