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UCHL5 binds Rpn13C. 31 / 31
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"Preliminary size exclusion chromatography (SEC) studies of Uch37 and its complex with Rpn13C revealed that at a concentration of approximately 10 mg/mL, Uch37 had a smaller retention volume than the Uch37 and Rpn13C complex."
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"This finding was unexpected and provided the first indication that the oligomerization of Uch37 might be different between Uch37 alone and the Uch37 and Rpn13C complex."
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"In sharp contrast, the elution volume of the Uch37 and Rpn13C complex was independent of the concentration."
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"The exact oligomerization state of Uch37 and Rpn13C complex could not be determined from the SEC profile because the theoretical MW of the complex calculated from its primary amino acid sequence is approximately 52.6 kDa."
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"We performed analytical ultracentrifugation (AUC) analysis to further confirm the MW and to obtain the oligomerization state of the Uch37 and Rpn13C complex."
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"This MW, which is close to the theoretical MW (52.6 kDa) of the Uch37 and Rpn13C complex, suggested that the Uch37 and Rpn13C complex existed as a heterodimer in solution."
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"A frictional ratio of 1.577 (f/f0> 1.2) indicated that the Uch37 and Rpn13C complex adopted an elongated shape in solution."
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"Figure XREF_FIG presents the SAXS data collected for different concentrations of Uch37 and the Uch37 and Rpn13C complex, and Table XREF_TABLE lists the parameters derived from those curves."
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"The Kratky plots of both Uch37 and the Uch37 and Rpn13C complex exhibited a clear peak with plateaus."
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"This finding suggested that both Uch37 and the Uch37 and Rpn13C complex were well folded and contained some flexible regions."
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"The MWs calculated from the SAXS P (r) functions for Uch37 and Uch37 and Rpn13C complex were approximately 117.7 and 64.6 kDa, respectively."
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"When compared to the theoretical MWs (Uch37, 37 kDa; Uch37 and Rpn13C complex, 52.6 kDa), the SAXS based MW estimations further highlighted the ability of Uch37 to form oligomers, although the Uch37 and Rpn13C complex might exist as a heterodimer."
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"A moderate increase in the I 0 implied that Rpn13C interacted with the oligomerized Uch37 [(Uch37) n], forming a larger intermediate (Uch37) n and Rpn13C complex."
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"To confirm that the interaction of Rpn13C with Uch37 was specific, we performed surface plasmon resonance (SPR) experiments and measured the affinity between Rpn13C and Uch37."
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"Uch37 bound to Rpn13C tightly with a low dissociation constant (K d = 5.35 nmol/L, Fig."
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"Interestingly, 4 mol/L NaCl could not reverse the interaction between Uch37 and Rpn13C, indicating a role for hydrophobic interactions in the formation of the complex."
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"A moderate increase in the I 0 implied that Rpn13C interacted with the oligomerized Uch37 [(Uch37)n], forming a larger intermediate (Uch37)n/Rpn13C complex."
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"Reconstruction of the Uch37 and Rpn13C complex structure by SAXS."
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"To visualize the interaction between Uch37 and Rpn13C, we employed ab initio and rigid body modeling methods to construct the Uch37 and Rpn13C complex from the SAXS data."
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"Ten individual GASBOR calculations without symmetry restraint (P1) were performed to construct a model for the Uch37 and Rpn13C complex."
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"The resultant model of the Uch37 and Rpn13C complex had an acceptable chi value of 1.44 +/- 0.21."
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"The consensus shape of the Uch37 and Rpn13C complex mainly contains two parts -- Uch37 (left part) and Rpn13C (right part)."
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"Furthermore, the atomic model of the Uch37 and Rpn13C complex generated by MASSHA could fit the ab initio low-resolution shape well."
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"In this SAXS based Uch37 and Rpn13C complex model, the Uch37 binding surface of Rpn13C reported by Chen et al. was facing toward the Uch37 molecule."
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"Although the overall SAXS based Uch37 and Rpn13C complex model is in agreement with the model reported by Chen et al., the distance between the Uch37 binding surface of Rpn13 and the Uch37 molecule is slightly larger."
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"This result is consistent with the inference derived from the biophysical characterization of the Uch37 and Rpn13C complex that Rpn13 activates Uch37 through its interaction with the C-terminal domain of Uch37, particularly the KEKE motif."
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"To confirm that the interaction of Rpn13C with Uch37 was specific, we performed surface plasmon resonance (SPR) experiments and measured the affinity between Rpn13C and Uch37."
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"Uch37 bound to Rpn13C tightly with a low dissociation constant ( K d = 5.35 nmol/L, Fig.  xref D)."
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"Interestingly, 4 mol/L NaCl could not reverse the interaction between Uch37 and Rpn13C, indicating a role for hydrophobic interactions in the formation of the complex (Fig.  xref E)."
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"To visualize the interaction between Uch37 and Rpn13C, we employed ab initio and rigid body modeling methods to construct the Uch37-Rpn13C complex from the SAXS data (Fig.  xref )."
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"In the presence of Rpn13C, Uch37 formed a 1:1 complex with Rpn13C. Such a configuration of Uch37 in the presence of Rpn13C readily cleaved the substrate Ub-AMC (Fig.  xref C)."
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