Solution Structure of the Immunodominant Region of Protein G of Bovine Respiratory Syncytial Virus

Protein G Of Bovine Respiratory Syncytial Virus

ABSTRACT

The three-dimensional solution structure of the immunodominant central conserved region of the attachment protein G (BRSV-G) of bovine respiratory syncytial virus has been determined by nuclear magnetic resonance (NMR) spectroscopy. In the 32-residue peptide studied, 19 residues form a small rigid core composed of two short helices, connected by a type I' turn, and linked by two disulfide bridges. This unique fold is among the smallest stable tertiary structures known and could therefore serve as an ideal building block for the design of de novo proteins and as a test case for modeling studies. A characteristic hydrophobic pocket, lined by conserved residues lies at the surface of the peptide and may play a role in receptor binding. This work provides a structural basis for further peptide vaccine development against the severe diseases associated with the respiratory syncytial viruses both in cattle and man.

Bovine respiratory syncytial virus (BRSV) accounts for a high proportion of morbidity and mortality in cattle (Stott & Taylor, 1985; Baker et al., 1986) resulting in considerable economic losses. Furthermore, human RSV (HRSV) is a major cause of serious respiratory infections including bronchiolitis and pneumonia in infants and young children: these conditions are responsible for almost 100,000 hospitalizations yearly in the USA alone (Heilman, 1990). The development of efficacious vaccines against HRSV infections has a very high priority according to the World Health Organization (World Health Organization, 1995), and has lately received much attention (Hall, 1994; McIntosh & Chanock, 1990).

Respiratory syncytial virus carries two glycoproteins on its surface: the fusion protein F and the attachment protein G. The latter is an integral membrane protein and is functionally distinct from other viral attachment proteins. The focus of the present study is the small conserved hydrophobic domain (Figure 1) that is located in the ectodomain and is presumably flanked in the intact protein G by two extended mucin-like regions (Langedijk et al., 1996b). This domain contains four conserved cysteines which form two disulfide bridges. Peptides based on this domain were found to be immunodominant in protein G of both HRSV (Norrby et al., 1987; Ekerlind-Stopner et al., 1990) and BRSV (Langedijk et al., 1996b); they can be used as antigens in highly specific and sensitive immunoassays (Langedijk et al., 1996a) and have conferred protection against HRSV challenge in mice (Trudel et al., 1991).

We have studied a 32-residue synthetic peptide representing the conserved central region of protein G of BRSV using two-dimensional (2D) NMR, distance geometry (DG), and restrained molecular dynamics (RMD).

Doreleijers, J.F., Langedijk, J.P.M., Herd, K., Boelens, R., Rullmann, J.A.C., Schaaper, W. M., van Oirschot, J.T., and Kaptein, R., "Solution Structure of the Immunodominant Region of Protein G of Bovine Respiratory Syncytial Virus," Biochemistry 35, 14684-14688 (1996).

Studied peptide was 32 AA in size. The NMR models of the 19 AA forming the structured part (171-189) are submitted to the PDB.

Solution Nmr Structure Of The Immunodominant Region Of Protein G Of Bovine Respiratory Syncytial Virus, 48 Structures

G protein [Bovine respiratory syncytial virus]

glycoprotein, partial [Bovine respiratory syncytial virus]

RecName: Full=Major surface glycoprotein G; AltName: Full=Attachment glycoprotein G; AltName: Full=Membrane-bound glycoprotein;