| dc.creator |
Arrondo, José Luis R. |
|
| dc.creator |
Echabe, Izaskun |
|
| dc.creator |
Iloro, Ibon |
|
| dc.creator |
Hernando, Miguel Ángel |
|
| dc.creator |
Cruz, Fernando de la |
|
| dc.creator |
Goñi, Félix M. |
|
| dc.date |
2008-04-14T09:05:25Z |
|
| dc.date |
2008-04-14T09:05:25Z |
|
| dc.date |
2003-07 |
|
| dc.date.accessioned |
2017-01-31T01:02:12Z |
|
| dc.date.available |
2017-01-31T01:02:12Z |
|
| dc.identifier |
Journal of Bacteriology 185(14): 4226–4232 (2003) |
|
| dc.identifier |
1098-5530 |
|
| dc.identifier |
http://hdl.handle.net/10261/3557 |
|
| dc.identifier |
10.1128/JB.185.14.4226-4232.2003 |
|
| dc.identifier.uri |
http://dspace.mediu.edu.my:8181/xmlui/handle/10261/3557 |
|
| dc.description |
The TrwC protein is the relaxase-helicase responsible for the initiation and termination reactions of DNA
processing during plasmid R388 conjugation. The TrwC-N275 fragment comprises the 275-amino-acid Nterminal
domain of the protein that contains the DNA cleavage and strand transfer activities (the relaxase
domain). It can be easily purified by keeping a cell lysate at 90°C for 10 min. Infrared spectroscopy shows that
this domain has a predominantly / structure with some amount of unordered structure. Fast heating and
cooling does not change the secondary structure, whereas slow heating produces two bands in the infrared
spectrum characteristic of protein aggregation. The denaturation temperature is increased in the protein after
the fast-heating thermal shock. Two-dimensional infrared correlation spectroscopy shows that thermal unfolding
is a very cooperative two-state process without any appreciable steps prior to aggregation. After
aggregation, the -helix percentage is not altered and -helix signal does not show in the correlation maps,
meaning that the helices are not affected by heating. The results indicate that the domain has an -helix core
resistant to temperature and responsible for folding after fast heating and an outer layer of -sheet and
unordered structure that aggregates under slow heating. The combination of a compact core and a flexible
outer layer could be related to the structural requirements of DNA-protein binding. |
|
| dc.description |
This work was supported by grants BMC2002-01438 from the Spanish
Ministry of Science and Technology and 9/UPV00042.310-13552
from the Universidad del País Vasco. F.D.L.C.’s work was supported
by grant PB98-1106 from the Spanish Ministry of Science and Technology. |
|
| dc.description |
Peer reviewed |
|
| dc.format |
768293 bytes |
|
| dc.format |
application/pdf |
|
| dc.language |
eng |
|
| dc.publisher |
American Society for Microbiology |
|
| dc.relation |
http://dx.doi.org/10.1128/JB.185.14.4226–4232.2003 |
|
| dc.rights |
closedAccess |
|
| dc.title |
A Bacterial TrwC Relaxase Domain Contains a Thermally Stable α-Helical Core |
|
| dc.type |
Artículo |
|