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. 2011 Sep 20;286(45):38939–38947. doi: 10.1074/jbc.M111.291377

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — The N-terminal 30-kDa fragment of fibronectin is sufficient for MAM7 binding. A, fibronectin subunit (220 kDa) consisting of type I, type II, and type III repeats. Each subunit contains an N-terminal region (I_1–5, 30-kDa fragment) required for fibrin and heparin binding (HBD), followed by a 45-kDa collagen binding domain (CBD, I₆, II_1–2, I_7–9). MAM7 contains an N-terminal transmembrane region, which was excluded from recombinant proteins, followed by seven consecutive mce domains. GST-MAM7 and MAM1 constructs used for pulldowns are depicted. Pulldown assays with GST-MAM7 and intact Fn (B), 30-kDa Fn subunit (Fn30kDa; C) or 45-kDa Fn subunit (Fn45kDa; C) or GST-MAM1 and 30-kDa Fn subunit (Fn30kDa; E). Heparin (hep) inhibits attachment of BL21-MAM7 to host cells (F). M7, BL21-MAM7; M7Δ, BL21-MAM7ΔTM. L, load; F, flow through; E, eluate; *, Fn30kDa resulting from breakdown of Fn.