Time filter

Source Type

Teaneck, NJ, United States

Rashin A.A.,BioChemComp Inc. | Rashin A.A.,Iowa State University | Jernigan R.L.,Iowa State University
Biochemistry | Year: 2016

Only α1 and α2 domains of the α-chain of the major histocompatibility complex (MHC) directly bind peptide antigens (Ag-s) and the T-cell receptor (TCR). Significant plasticity was found in the TCR but only minor in (α1 + α2). The α3-domain position variation was noted only in connection to its binding the coreceptor CD8. We apply our methods for identifying functional conformational changes in proteins to a systematic study of similarities between 43 X-ray structures of the entire α chains of MHC-I HLA-A2. Out of 903 different αHLA-A2 pairs 203 show similarities within the earlier determined uncertainty threshold and unexpectedly form three similarity clusters (SCs) with all/most structures in a cluster similar within the uncertainty threshold. Pairs from different SCs always differ above the threshold, mainly due to variations in the α3 position/structure. All structures in SC3 cannot bind the CD8 coreceptor. Strong hydrogen bonds between (α1 + α2) and α3 differ between SC1 and SC2 but are nearly invariant within each SC. Small conformational changes in the (α1 + α2), caused by Ag-s differences, act as an α3 "allosteric switch" between SC2 and SC1. Binding of CD8 to SC2-HLA-A2 (Tax-type Ag-s) changes it to SC1-HLA-A2 (HuD-type Ag-s). HuD binding to HLA-A2 is much less stable than Tax binding. CD8-liganded HLA-A2 preference for binding HuD suggests that CD8-HLA-A2 may present a weakly binding peptide for TCR recognition, supporting the hypothesis that CD8 increases TCR avidity to weak Ag-s. Other HLA-A2 functions may involve α3. TCR-A6-liganded-Tax-type-HLA-A2s form two small clusters, similar to either A6-liganded-HuD or A6-liganded-native-Tax HLA-A2s. © 2015 American Chemical Society. Source

Rashin A.A.,BioChemComp Inc. | Rashin A.A.,Iowa State University | Rashin A.H.L.,BioChemComp Inc. | Rashin A.H.L.,Rutgers University | Jernigan R.L.,Iowa State University
Biochemistry | Year: 2010

It was found that the variety of function-related conformational changes ("movements") in proteins is beyond the earlier simple classifications. Here we offer biochemists a more comprehensive, transparent, and easy-to-use approach allowing a detailed and accurate interpretation of such conformational changes. It makes possible a more multifaceted characterization of protein flexibility via identification of rigidly and nonrigidly repositioned fragments, stable and nonstable fragments, and domain and nondomain repositioning. "Coordinate uncertainty thresholds" derived from computed differences between independently determined coordinates of the same molecules are used as the criteria for conformational identity. "Identical" rigid substructures are localized in the distance difference matrices (DDMs). A sequence of simple transformations determines whether a structural change occurs by rigid-body movements of fragments or largely through non-rigid-body deformations. We estimate the stability of protein fragments and compare stable and rigidly moving fragments. The motions computed with the coarse-grained elastic networks are also compared to those of their DDM analogues. We study and suggest a classification for 17 structural pairs, differing in their functional states. For five of the 17 proteins, conformational change cannot be accomplished by rigid-body transformations and requires significant non-rigid-body deformations. Stable fragments rarely coincide with rigidly moving fragments and often disagree with the CATH identifications of domains. Almost all monomeric apo chains, containing stable fragments and/or domains, indicate instability of the entire molecule, suggesting the importance of fragments and domains motions prior to stabilization by substrate binding or crystallization. Notably, kinases exhibit the greatest extent of nonrigidity among the proteins investigated. © 2010 American Chemical Society. Source

Rashin A.A.,BioChemComp Inc. | Rashin A.A.,Iowa State University | Domagalski M.J.,University of Virginia | Zimmermann M.T.,Iowa State University | And 4 more authors.
Acta Crystallographica Section D: Biological Crystallography | Year: 2014

Validation of general ideas about the origins of conformational differences in proteins is critical in order to arrive at meaningful functional insights. Here, principal component analysis (PCA) and distance difference matrices are used to validate some such ideas about the conformational differences between 291 myoglobin structures from sperm whale, horse and pig. Almost all of the horse and pig structures form compact PCA clusters with only minor coordinate differences and outliers that are easily explained. The 222 whale structures form a few dense clusters with multiple outliers. A few whale outliers with a prominent distortion of the GH loop are very similar to the cluster of horse structures, which all have a similar GH-loop distortion apparently owing to intermolecular crystal lattice hydrogen bonds to the GH loop from residues near the distal histidine His64. The variations of the GH-loop coordinates in the whale structures are likely to be owing to the observed alternative intermolecular crystal lattice bond, with the change to the GH loop distorting bonds correlated with the binding of specific 'unusual' ligands. Such an alternative intermolecular bond is not observed in horse myoglobins, obliterating any correlation with the ligands. Intermolecular bonds do not usually cause significant coordinate differences and cannot be validated as their universal cause. Most of the native-like whale myoglobin structure outliers can be correlated with a few specific factors. However, these factors do not always lead to coordinate differences beyond the previously determined uncertainty thresholds. The binding of unusual ligands by myoglobin, leading to crystal-induced distortions, suggests that some of the conformational differences between the apo and holo structures might not be 'functionally important' but rather artifacts caused by the binding of 'unusual' substrate analogs. The causes of P6 symmetry in myoglobin crystals and the relationship between crystal and solution structures are also discussed. © 2014 International Union of Crystallography. Source

Discover hidden collaborations