2005
DOI: 10.1074/jbc.m410206200
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The High Resolution Crystal Structure of a Native Thermostable Serpin Reveals the Complex Mechanism Underpinning the Stressed to Relaxed Transition

Abstract: Serpins fold into a native metastable state and utilize a complex conformational change to inhibit target proteases. An undesirable result of this conformational flexibility is that most inhibitory serpins are heat sensitive, forming inactive polymers at elevated temperatures. However, the prokaryote serpin, thermopin, from Thermobifida fusca is able to function in a heated environment. We have determined the 1.8 Å x-ray crystal structure of thermopin in the native, inhibitory conformation. A structural compar… Show more

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Cited by 30 publications
(32 citation statements)
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“…2) in which the RCL is completely exposed. For example, the main chain trace within this region of CBG is identical to those of uncleaved human AAT (32)(33)(34), thermopin (35), and some noninhibitory serpins such as ovalbumin (36) and maspin (37). Interestingly, the conformation of the RCL observed in CBG also resembles that of other ligand-bound serpins, such as the complex of antithrombin with a pentasaccharide ligand (21, 38) (Fig.…”
Section: Resultsmentioning
confidence: 72%
“…2) in which the RCL is completely exposed. For example, the main chain trace within this region of CBG is identical to those of uncleaved human AAT (32)(33)(34), thermopin (35), and some noninhibitory serpins such as ovalbumin (36) and maspin (37). Interestingly, the conformation of the RCL observed in CBG also resembles that of other ligand-bound serpins, such as the complex of antithrombin with a pentasaccharide ligand (21, 38) (Fig.…”
Section: Resultsmentioning
confidence: 72%
“…Given the prediction that the side chain of Glu-90 was solvent-exposed, we did not anticipate that this substitution would alter the overall structure of the native serpin fold. However, Glu-90 was located close to the shutter region, and as demonstrated with the Thermobifida fusca serpin, thermopin, even minor substitutions in the shutter can lead to a profound loss of inhibitory activity (32). Residue Pro-303 was predicted to be located at the C-terminal end of hI (Fig.…”
Section: Discussionmentioning
confidence: 99%
“…The shutter region, which is centered on the top of helix B and includes parts of helix D, contains important determinants for the mobility of ␤-sheet A. This region is highly sensitive to change, since mutations in helix B abolish the inhibitory activity of thermopin without affecting its heat denaturation profile (32), and mutations in helix D cause spontaneous conformational changes in SERPINC1/antithrombin III (33). Mutations in helix I are less common, but an S349P mutation of SERPINC1 leads to increased thrombosis and a type 2 deficiency (normal amounts of circulating protein but decreased activity) (34).…”
mentioning
confidence: 99%
“…Structural studies reveal that thermopin possesses an extreme C-terminal sequence that folds across the front of the molecule and interacts with the top of sheet A (Fig. 3A) (21,24). Biophysical studies show that the C-terminal sequence plays no detectable role in influencing the stability of the native fold but instead appears to be important for stabilizing a folding intermediate.…”
Section: Serpins From Thermophilic Organisms Provide New Insights Intmentioning
confidence: 99%