The nature of the supramolecular complex between fibrillar collagen and collagen-binding proteins (CBPs) has hindered detailed X-ray and NMR analyses of the ligand-recognition mechanism at atomic resolution because of the lack of appropriate approaches for studying large heterogeneous supramolecular complexes. Recently, we proposed an NMR method, termed transferred cross-saturation (TCS), that enables the rigorous identification of contact residues in a huge protein complex. Here we used TCS to study the supramolecular complex between the A3 domain of von Willebrand factor and fibrillar collagen, which allowed the successful determination of the ligand-binding site of the A3 domain. The binding site of the A3 domain was located at its hydrophobic 'front' surface and was completely different from that of the I domain from the a2 subunit of integrin (alpha2-I domain), which was reported to be the hydrophilic 'top' surface of alpha2-I, although the A3 domain and the alpha2-I domain share a similar fold and possess the identical function of collagen binding.
A polyclonal antibody (anti-PepTl/C) was raised fine, and the predicted protein, termed PepT1 [11], consists of against the rabbit intestinal H+-coupled oligopeptide transporter, 707 amino acids and has 12 membrane-spanning domains. On PepT1. Anti-PepTl/C detected 70-80-kDa protein in crude the basis of in vitro translation, the apparent molecular mass membranes obtained from rabbit duodenum, jejunum and ileum, of the product is 71 kDa.
PepT1 was localized in the brush-border of the absorptiveIn the present study, we raised a polyclonal antibody epithelial cells by subcellular fractionation of membranes on a against a synthetic peptide corresponding to a fragment of sucrose density gradient and by immunohistochemistry using antibiotics [8,25,26]. So, the ability of PepT1 to transport orally active [Mactam antibiotics was also examined by expression of the mRNA in Xenopus oocytes.
Human growth hormone (hGH) induces dimerization of its binding protein (hGHbp). hGH binds to the first hGHbp (bp1) on site 1, and then the hGH-bp1 heterodimer complex binds to the second hGHbp (bp2) on site 2. Although the interactions of hGH and hGHbps have been studied from different viewpoints, few studies from a dynamic viewpoint have been reported. Especially, since in the SCOP domain database hGHbp is classified as two clear immunoglobulin-like domains, it is of interest to understand how hGH interacts with the hGHbp domains. Therefore, we carried out normal mode analysis (NMA) of free hGH, free bp1, free bp2, and the hGH-bp1 heterodimer complex, as well as the hGH-bp1-bp2 ternary complex to investigate how the dynamics of the proteins change before and after forming the complexes. NMA showed that the domain motion between the N-terminal and the C-terminal domains of free bp1 markedly decreased after binding to hGH, and that the domain motion of bp2 decreased similarly after binding to the hGH-bp1 heterodimer complex. The present study demonstrates that hGH regulates the inter-domain motions of both hGHbps.
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