Cloned (Comment) | Organism |
---|---|
cDNAs of full-length Rpp20 and Rpp25 cloned into pCR4-TOPO. Full-length Rpp25 (encompassing residues 1-199) subcloned with an N-terminal hexahistidine tag into a pPROEX-HTb expression vector using Nco1/Not1 restriction sites. Full-length Rpp20 (residues 1-140) subcloned into a pET-30 expression vector. The pET-30 vector modified to bear a TEV cleavage site to remove the N-terminal hexahistidine tag. Mutant Rpp20(35-140) amplified from pET-30 Rpp20 to introduce 5' Asc1 and 3' Not1 sites and then ligated into a modified version of pET-15b vector with an Asc1 site inserted directly after the his tag. Fragments Rpp20, mutant Rpp20(16-140), mutant Rpp25(25-170) and co-expressed Rpp20/Rpp25 (in which Rpp20 bears the his tag) subcloned in pETDuet-1 vector with an N-terminal hexahistidine tag. Mutant Rpp25(25-170) also subcloned in a pCDFDuet-1 vector also with an N-terminal hexahistidine tag. Rpp25, Rpp20 (from pET-30 vector) and mutant Rpp20(35-140) expressed in Escherichia coli strain BL21(DE3)pLysS. All the other proteins subcloned into pETDuet-1 or pCDFDuet-1 vectors expressed in Escherichia coli KRX strain | Homo sapiens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
13200 | - |
2 * 16000, full-length Rpp20, gel filtration. 2 * 14100, Rpp20(16-140), gel filtration. 2 * 13200, HisRpp20(35-140), gel filtration | Homo sapiens |
14100 | - |
2 * 16000, full-length Rpp20, gel filtration. 2 * 14100, Rpp20(16-140), gel filtration. 2 * 13200, HisRpp20(35-140), gel filtration | Homo sapiens |
15400 | - |
mutant Rpp25(25-170), gel filtration | Homo sapiens |
16000 | - |
2 * 16000, full-length Rpp20, gel filtration. 2 * 14100, Rpp20(16-140), gel filtration. 2 * 13200, HisRpp20(35-140), gel filtration | Homo sapiens |
20600 | - |
mutant Rpp25, gel filtration | Homo sapiens |
20700 | - |
mutant Rpp25(25-170), sequence analysis | Homo sapiens |
26400 | - |
mutant HisRpp20(35-140), gel filtration | Homo sapiens |
28100 | - |
mutant Rpp20(16-140), gel filtration | Homo sapiens |
28600 | - |
mutant Rpp20(35-140)/Rpp25(25-170), gel filtration | Homo sapiens |
31100 | - |
mutant Rpp20(35-140)/Rpp25(25-170), sequence analysis | Homo sapiens |
31500 | - |
mutant Rpp20/Rpp25(25-170), gel filtration | Homo sapiens |
32100 | - |
full-length Rpp20, gel filtration | Homo sapiens |
36600 | - |
mutant Rpp20/Rpp25, gel filtration | Homo sapiens |
39200 | - |
mutant Rpp25, sequence analysis | Homo sapiens |
39900 | - |
mutant HisRpp20(35-140), sequence analysis | Homo sapiens |
41100 | - |
mutant Rpp20/Rpp25(25-170), sequence analysis | Homo sapiens |
44400 | - |
mutant [Rpp20(35-140)-Rpp25(25-170)]/P3 RNA, gel filtration | Homo sapiens |
45300 | - |
mutant Rpp20(16-140), sequence analysis | Homo sapiens |
47000 | - |
full-length Rpp20, sequence analysis | Homo sapiens |
52500 | - |
mutant [Rpp20-Rpp25]/P3 RNA, gel filtration | Homo sapiens |
53800 | - |
mutant Rpp20/Rpp25, sequence analysis | Homo sapiens |
56400 | - |
mutant [Rpp20(35-140)-Rpp25(25-170)]/P3 RNA, sequence analysis | Homo sapiens |
71400 | - |
mutant [Rpp20-Rpp25]/P3 RNA, sequence analysis | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Source Tissue | Comment | Organism | Textmining |
---|
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Rpp20 and Rpp25 interact with the P3 arm of RNase MRP RNA in a highly synergic fashion. Rpp20 and Rpp25 interact with the P3 RNA as a heterodimer, which is formed prior to RNA binding | Homo sapiens | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterodimer | Rpp20 and Rpp25, ITC-200 microcalorimeter experiments | Homo sapiens |
homodimer | 2 * 16000, full-length Rpp20, gel filtration. 2 * 14100, Rpp20(16-140), gel filtration. 2 * 13200, HisRpp20(35-140), gel filtration | Homo sapiens |
monomer | Rpp25 in solution | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
RNase P | - |
Homo sapiens |
RNase P/MRP | - |
Homo sapiens |
Rpp20 | - |
Homo sapiens |
Rpp25 | - |
Homo sapiens |
General Information | Comment | Organism |
---|---|---|
malfunction | in both deletion mutants Rpp20(16-140) and Rpp20(35-140) the global thermodynamics of the interaction with Rpp25 is seemingly unaffected. Thermodynamic signature of the association is also fully preserved between the Rpp25(25-170) mutant and all available versions of Rpp20. Regions within the mutants Rpp20(35-140) and Rpp25(25-170) are sufficient for mutual interaction, thus this recognition can be mediated largely, if not exclusively, by the Alba-type core domains | Homo sapiens |
physiological function | strong interaction between Rpp25 and Rpp20. Rpp20 and Rpp25 interact with the P3 arm of RNase MRP RNA in a highly synergic fashion. Rpp20 and Rpp25 interact with the P3 RNA as a heterodimer, which is formed prior to RNA binding. Association between Rpp20 and Rpp25 has no detectable influence on their secondary/tertiary structure. The association reaction results in a large loss of solvent-accessible area. N- and C-terminal regions of Rpp25 and the N-terminal tail of Rpp20 are not involved in mutual recognition | Homo sapiens |