Novel fold and capsid-binding properties of the lambda-phage display platform protein gpD.
The crystal structure of gpD, the capsid-stabilizing protein of bacteriophage lambda, was solved at 1.1 A resolution. Data were obtained from twinned crystals in space group P21 and refined with anisotropic temperature factors to an R-factor of 0.098 (Rfree = 0. 132). GpD (109 residues) has a novel fold with an unusually low content of regular secondary structure. Noncrystallographic trimers with substantial intersubunit interfaces were observed. The C-termini are well ordered and located on one side of the trimer, relatively far from its three-fold axis. The N-termini are disordered up to Ser 15, which is close to the three-fold axis and on the same side as the C-termini. A density map of the icosahedral viral capsid at 15 A resolution, obtained by cryo-electron microscopy and image reconstruction, reveals gpD trimers, seemingly indistinguishable from the ones seen in the crystals, at all three-fold sites. The map further reveals that the side of the trimer that binds to the capsid is the side on which both termini reside. Despite this orientation of the gpD trimer, fusion proteins connected by linker peptides to either terminus bind to the capsid, allowing protein and peptide display.
Yang
F
F
Macromolecular Crystallography Laboratory, Program in Structural Biology, National Cancer Institute, Frederick Cancer Research and Development Center, Frederick, Maryland 21702, USA.
Forrer
P
P
Dauter
Z
Z
Conway
J F
JF
Cheng
N
N
Cerritelli
M E
ME
Steven
A C
AC
Plückthun
A
A
Wlodawer
A
A
D000595:Amino Acid Sequence; D010582:Bacteriophage lambda / Q000737:chemistry*; D002213:Capsid / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D036022:Capsid Proteins; D002850:Chromatography, Gel; D002942:Circular Dichroism; D020285:Cryoelectron Microscopy; D003460:Crystallization; D018360:Crystallography, X-Ray; D006023:Glycoproteins / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D006860:Hydrogen Bonding; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D019151:Peptide Library; D011485:Protein Binding; D017510:Protein Folding; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D012996:Solutions; D013696:Temperature; D014764:Viral Proteins / Q000378:metabolism / Q000648:ultrastructure
D036022:Capsid Proteins; D006023:Glycoproteins; D019151:Peptide Library; D012996:Solutions; D014764:Viral Proteins; C406073:gpD protein, lambda phage