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Specific interactions of serpins in their native forms attenuate their conformational transitions
Author(s) -
Na YuRan,
Im Hana
Publication year - 2007
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.072838107
Subject(s) - serpin , beta sheet , biophysics , chemistry , protein structure , förster resonance energy transfer , serine protease , protein folding , plasminogen activator inhibitor 1 , protein secondary structure , transition (genetics) , conformational change , circular dichroism , plasminogen activator , stereochemistry , biochemistry , biology , protease , fluorescence , genetics , enzyme , gene , physics , quantum mechanics
Plasminogen activator inhibitor‐1 (PAI‐1) belongs to the serine protease inhibitor (serpin) protein superfamily. Serpins are unique in that their native forms are not the most thermodynamically stable conformation; instead, a more stable, latent conformation exists. During the transition to the latent form, the first strand of β‐sheet C (s1C) in the serpin is peeled away from the β‐sheet, and the reactive center loop (RCL) is inserted into β‐sheet A, rendering the serpin inactive. To elucidate the contribution of specific interactions in the metastable native form to the latency transition, we examined the effect of mutations at the s1C of PAI‐1, specifically in positions P4′ through P10′. Several mutations strengthened the interactions between these residues and the core protein, and slowed the transition of the protein from the metastable native form to the latent form. In particular, anchoring of the strand to the protein's hydrophobic core at the beginning (P4′ site) and center of the strand (P8′ site) greatly retarded the latency transition. Mutations that weakened the interactions at the s1C region facilitated the conformational conversion of the protein to the latent form. PAI‐1's overall structural stability was largely unchanged by the mutations, as evaluated by urea‐induced equilibrium unfolding monitored via fluorescence emission. Therefore, the mutations likely exerted their effects by modulating the height of the energy barrier from the native to the latent form. Our results show that interactions found only in the metastable native form of serpins are important structural features that attenuate folding of the proteins into their latent forms.