To investigate the contribution of the folding cores to the thermodynamic stability of RNases H, we used rational design to create two chimeras composed of parts of a thermophilic and a mesophilic RNase H. Each chimera combines the folding core from one parent protein and the remaining parts of the other. Both chimeras form active, well-folded RNases H. Stability curves, based on CD-monitored chemical denaturations, show that the chimera with the thermophilic core is more stable, has a higher midpoint of thermal denaturation, and a lower change in heat capacity (DeltaCp) upon unfolding than the chimera with the mesophilic core. A possible explanation for the low DeltaCp of both the parent thermophilic RNase H and the chimera with the thermophilic core is the residual structure in the denatured state. On the basis of the studied parameters, the chimera with the thermophilic core resembles a true thermophilic protein. Our results suggest that the folding core plays an essential role in conferring thermodynamic parameters to RNases H. Study holds ProTherm entries: 13061, 13062, 13063, 13064, 13065, 13066 Extra Details: thermodynamic stability of proteins; chimera; stability curves; heat capacity; folding core; ribonuclease H
ID: RMCVWfDR
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:44 p.m.
Version: 1
Colors: | D | E | R | H | K | S | T | N | Q | A | V | I | L | M | F | Y | W | C | G | P |
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Percent Identity | Matching Chains | Protein | Accession | Entry Name |
---|---|---|---|---|
100.0 | Ribonuclease H | P29253 | RNH_THET8 | |
98.8 | Ribonuclease H | Q72IE1 | RNH_THET2 | |
100.0 | Ribonuclease HI | A7ZHV1 | RNH_ECO24 | |
100.0 | Ribonuclease HI | B7MBJ0 | RNH_ECO45 | |
100.0 | Ribonuclease HI | P0A7Y6 | RNH_ECO57 | |
100.0 | Ribonuclease HI | B5Z0I8 | RNH_ECO5E | |
100.0 | Ribonuclease HI | B7NKW4 | RNH_ECO7I | |
100.0 | Ribonuclease HI | B7MQ23 | RNH_ECO81 | |
100.0 | Ribonuclease HI | B7M213 | RNH_ECO8A | |
100.0 | Ribonuclease HI | C4ZRV1 | RNH_ECOBW | |
100.0 | Ribonuclease HI | B1XD78 | RNH_ECODH | |
100.0 | Ribonuclease HI | P0A7Y5 | RNH_ECOL6 | |
100.0 | Ribonuclease HI | B1IPU4 | RNH_ECOLC | |
100.0 | Ribonuclease HI | P0A7Y4 | RNH_ECOLI | |
100.0 | Ribonuclease HI | B7N876 | RNH_ECOLU | |
100.0 | Ribonuclease HI | B6HZS7 | RNH_ECOSE | |
100.0 | Ribonuclease HI | B1LHM3 | RNH_ECOSM | |
100.0 | Ribonuclease HI | B7LW89 | RNH_ESCF3 | |
100.0 | Ribonuclease HI | B2U352 | RNH_SHIB3 | |
100.0 | Ribonuclease HI | Q325T2 | RNH_SHIBS | |
100.0 | Ribonuclease HI | Q32JP9 | RNH_SHIDS | |
100.0 | Ribonuclease HI | P0A7Y7 | RNH_SHIFL | |
100.0 | Ribonuclease HI | Q3Z5E9 | RNH_SHISS | |
99.4 | Ribonuclease HI | B7UJB0 | RNH_ECO27 | |
99.4 | Ribonuclease HI | B7LHC0 | RNH_ECO55 | |
99.4 | Ribonuclease HI | A7ZWF6 | RNH_ECOHS | |
99.4 | Ribonuclease HI | Q0TLC3 | RNH_ECOL5 | |
93.5 | Ribonuclease HI | A8AKR0 | RNH_CITK8 | |
93.5 | Ribonuclease HI | B5F8X2 | RNH_SALA4 | |
93.5 | Ribonuclease HI | A9MPF1 | RNH_SALAR | |
93.5 | Ribonuclease HI | Q57SZ6 | RNH_SALCH | |
93.5 | Ribonuclease HI | B5FJ58 | RNH_SALDC | |
93.5 | Ribonuclease HI | B5R449 | RNH_SALEP | |
93.5 | Ribonuclease HI | B5R5L3 | RNH_SALG2 | |
93.5 | Ribonuclease HI | B4TK85 | RNH_SALHS | |
93.5 | Ribonuclease HI | B4SV39 | RNH_SALNS | |
93.5 | Ribonuclease HI | Q5PFD8 | RNH_SALPA | |
93.5 | Ribonuclease HI | A9MZ19 | RNH_SALPB | |
93.5 | Ribonuclease HI | B5BDW5 | RNH_SALPK | |
93.5 | Ribonuclease HI | B4TYH0 | RNH_SALSV | |
93.5 | Ribonuclease HI | P0A2C0 | RNH_SALTI | |
93.5 | Ribonuclease HI | P0A2B9 | RNH_SALTY | |
92.9 | Ribonuclease HI | C0Q6N2 | RNH_SALPC | |
90.9 | Ribonuclease HI | B5Y1G2 | RNH_KLEP3 | |
90.9 | Ribonuclease HI | A6T512 | RNH_KLEP7 |