1iyb: Difference between revisions
New page: left|200px<br /><applet load="1iyb" size="450" color="white" frame="true" align="right" spinBox="true" caption="1iyb, resolution 1.50Å" /> '''Crystal Structure of... |
No edit summary |
||
| Line 1: | Line 1: | ||
[[Image:1iyb.gif|left|200px]]<br /><applet load="1iyb" size=" | [[Image:1iyb.gif|left|200px]]<br /><applet load="1iyb" size="350" color="white" frame="true" align="right" spinBox="true" | ||
caption="1iyb, resolution 1.50Å" /> | caption="1iyb, resolution 1.50Å" /> | ||
'''Crystal Structure of the Nicotiana glutinosa Ribonuclease NW'''<br /> | '''Crystal Structure of the Nicotiana glutinosa Ribonuclease NW'''<br /> | ||
==Overview== | ==Overview== | ||
Ribonuclease NW (RNase NW), the wound-inducible RNase in Nicotiana | Ribonuclease NW (RNase NW), the wound-inducible RNase in Nicotiana glutinosa leaves, preferentially cleaves guanylic acid. We expressed the cDNA encoding RNase NW in the methylotrophic yeast Pichia pastoris using the expression vector pPIC9K, and the resulting recombinant RNase NW (ryRNaseNW) secreted into medium was purified to apparent homogeneity using column chromatography. The crystal structure of ryRNase NW bound to 5'-GMP was determined at 1.5 A resolution by molecular replacement with tomato RNase LE as a search model. The RNase NW structurally belongs to the (alpha + beta) class of proteins, having eight helices (five alpha-helices and three 3(10) helices) and six beta-strands, and its structure is highly similar to those of other plant RNases, including a uridylic acid preferential RNase MC1 from bitter gourd seeds. The guanine ring of 5'-GMP lies in a hydrophobic pocket of the molecular surface composed of Tyr17, Tyr71, Ala80, Leu79, and Phe89: the guanine base is sandwiched between aromatic side chains of Tyr17 and Phe89. In addition, the guanine base is firmly stabilized by a network of hydrogen bonds of the side chains of Gln12 and Thr78, as well as of the main chain of Leu79. Therefore, Gln12, Tyr17, Thr78, Leu79, and Phe89 are responsible for recognition of the guanine base by RNase NW, findings which provide insight into the manner in which RNase NW preferentially cleaves guanylic acid. | ||
==About this Structure== | ==About this Structure== | ||
1IYB is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Nicotiana_glutinosa Nicotiana glutinosa] with 5GP as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http:// | 1IYB is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Nicotiana_glutinosa Nicotiana glutinosa] with <scene name='pdbligand=5GP:'>5GP</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1IYB OCA]. | ||
==Reference== | ==Reference== | ||
| Line 20: | Line 20: | ||
[[Category: ribonuclease]] | [[Category: ribonuclease]] | ||
''Page seeded by [http:// | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:16:55 2008'' | ||
Revision as of 14:17, 21 February 2008
|
Crystal Structure of the Nicotiana glutinosa Ribonuclease NW
OverviewOverview
Ribonuclease NW (RNase NW), the wound-inducible RNase in Nicotiana glutinosa leaves, preferentially cleaves guanylic acid. We expressed the cDNA encoding RNase NW in the methylotrophic yeast Pichia pastoris using the expression vector pPIC9K, and the resulting recombinant RNase NW (ryRNaseNW) secreted into medium was purified to apparent homogeneity using column chromatography. The crystal structure of ryRNase NW bound to 5'-GMP was determined at 1.5 A resolution by molecular replacement with tomato RNase LE as a search model. The RNase NW structurally belongs to the (alpha + beta) class of proteins, having eight helices (five alpha-helices and three 3(10) helices) and six beta-strands, and its structure is highly similar to those of other plant RNases, including a uridylic acid preferential RNase MC1 from bitter gourd seeds. The guanine ring of 5'-GMP lies in a hydrophobic pocket of the molecular surface composed of Tyr17, Tyr71, Ala80, Leu79, and Phe89: the guanine base is sandwiched between aromatic side chains of Tyr17 and Phe89. In addition, the guanine base is firmly stabilized by a network of hydrogen bonds of the side chains of Gln12 and Thr78, as well as of the main chain of Leu79. Therefore, Gln12, Tyr17, Thr78, Leu79, and Phe89 are responsible for recognition of the guanine base by RNase NW, findings which provide insight into the manner in which RNase NW preferentially cleaves guanylic acid.
About this StructureAbout this Structure
1IYB is a Single protein structure of sequence from Nicotiana glutinosa with as ligand. Full crystallographic information is available from OCA.
ReferenceReference
Guanine binding site of the Nicotiana glutinosa ribonuclease NW revealed by X-ray crystallography., Kawano S, Kakuta Y, Kimura M, Biochemistry. 2002 Dec 24;41(51):15195-202. PMID:12484757
Page seeded by OCA on Thu Feb 21 13:16:55 2008