Structure of the newly found green turtle egg-white ribonuclease.
Marine green turtle (Chelonia mydas) egg-white ribonuclease (GTRNase) was crystallized from 1.1 M ammonium sulfate pH 5.5 and 30% glycerol using the sitting-drop vapour-diffusion method. The structure of GTRNase has been solved at 1.60 A resolution by the molecular-replacement technique using a model based on the structure of RNase 5 (murine angiogenin) from Mus musculus (46% identity). The crystal belonged to the monoclinic space group C2, with unit-cell parameters a = 86.271, b = 34.174, c = 39.738 A, alpha = 90, beta = 102, gamma = 90 degrees . GTRNase consists of three helices and seven beta-strands and displays the alpha+beta folding topology typical of a member of the RNase A superfamily. Superposition of the C(alpha) coordinates of GTRNase and RNase A superfamily members indicates that the overall structure is highly similar to that of angiogenin or RNase 5 from M. musculus (PDB code 2bwl) and RNase A from Bos taurus (PDB code 2blz), with root-mean-square deviations of 3.9 and 2.0 A, respectively. The catalytic residues are conserved with respect to the RNase A superfamily. The three disulfide bridges observed in the reptilian enzymes are conserved in GTRNase, while one further disulfide bond is required for the structural stability of mammalian RNases. GTRNase is expressed in egg white and the fact that its sequence has the highest similarity to that of snapping turtle pancreatic RNase suggests that the GTRNase secreted from oviduct cells to form egg white is probably the product of the same gene as activated in pancreatic cells.
Katekaew
Somporn
S
Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen 42000, Thailand.
Kuaprasert
Buabarn
B
Torikata
Takao
T
Kakuta
Yoshimitsu
Y
Kimura
Makoto
M
Yoneda
Kazunari
K
Araki
Tomohiro
T
D000465:Algorithms; D000595:Amino Acid Sequence; D000818:Animals; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010063:Ovum / Q000201:enzymology*; D017434:Protein Structure, Tertiary; D012260:Ribonucleases / Q000737:chemistry*; D016415:Sequence Alignment; D040681:Structural Homology, Protein; D014426:Turtles / Q000378:metabolism*
D012260:Ribonucleases