Three-dimensional structure of human RNase 1 delta N7 at 1.9 A resolution.
Human pancreatic ribonuclease 1 (RNase 1) is considered to be the human counterpart of bovine pancreatic RNase A. Truncation of seven amino-acid residues from the amino-terminal sequence resulted in RNase 1 Delta N7, which has a reduced ribonucleolytic activity and a lower affinity for the human placental RNase inhibitor (PRI). This RNase 1 variant has been cloned, heterologously overexpressed, purified and crystallized. Its crystal structure has been determined and refined using data to 1.9 A resolution. The molecule displays the alpha + beta folding topology typical of members of the RNase A superfamily. The main distinct features found in RNase 1 Delta N7 are basically located in three loops affecting the fitting of the enzyme to the active site of subtilisin and the shape of the B2 subsite. These changes, taken with the lack of the catalytically active residue Lys7, may explain the reduced affinity of RNase 1 Delta N7 for PRI and the low ribonucleolytic activity of the protein when compared with the native enzyme.
Pous
J
J
Institut de Biologia Molecular de Barcelona, CID--CSIC, Jordi Girona 18--26, 08034 Barcelona, Spain.
Mallorquí-Fernández
G
G
Peracaula
R
R
Terzyan
S S
SS
Futami
J
J
Tada
H
H
Yamada
H
H
Seno
M
M
de Llorens
R
R
Gomis-Rüth
F X
FX
Coll
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D018360:Crystallography, X-Ray; D004722:Endoribonucleases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism; D004926:Escherichia coli; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D012259:Ribonuclease, Pancreatic / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion / Q000235:genetics*
D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D004722:Endoribonucleases; C046366:angiogenin; D012259:Ribonuclease, Pancreatic