IndraLab

Statements

P2 binds STAMBP. 28 / 28
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"The backbone and side-chain resonance assignments of UIM-SH3 bound to AMSH-P2 have been deposited to the BMRB under the accession number 30041."
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"The backbone resonance assignment of the UIM-SH3 construct in the free state has been obtained previously [24] , while the backbone assignment of UIM-SH3 in complex with AMSH-P2 has been obtained by m[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"Intermolecular NOEs between aliphatic/aromatic protons of UIM-SH3 and AMSH-P2 were obtained from 3D NOESY-HSQC (mixing time equal to 200 ms) and ( 15 N, 13 C)-filtered, 13 C-edited 3D NOESY-HSQC (mixi[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"In the case of the 15 N-UIM-SH3/AMSH-P2 complex or the 15 N-UIM-SH3 + AMSH-P2 + Lys63-Ub 2 complex, we used transverse relaxation periods of 10, 30, 50, 70 (twice), 90, 130, 170 (twice), 210, and 230 [MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"The whole AMSH-P2 peptide has been defined as fully flexible."
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"For AMSH-P2, the average pattern of CSPs did not exhibit any drastic change except for Tyr218, Asp219, and Ala244 ( Fig. 2 b)."
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"Additionally, the peptides containing half of the AMSH-P2 sequence (AMSH-P5 and P6) did not exhibit any significant CSPs and hence no significant binding with SH3 (see Fig. S12A and B)."
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"From the titration curves (Fig. S14B), we derived a dissociation constant of 563 ± 120.0 μM that is practically 200-fold greater compared to the binding of AMSH-P2 with SH3."
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"To address this question, we used the AMSH-P2 peptide and analyzed the CSPs of the UIM-SH3/Lys63-Ub 2 complex in the presence of AMSH-P2."
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"To an 15 N-UIM-SH3 sample saturated by Lys63-Ub 2 , we gradually added AMSH-P2 and monitored the CSPs in the corresponding 1 H, 15 N-HSQC spectrum."
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"In both cases, this result reflects an increase of the molecular mass by an average of ~ 2.7 ± 0.8 kDa, with respect to the SH3 domain of UIM-SH3 in the free state, which is hence close to the molecul[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
27725184
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"According to our CSP mapping, the binding interface resulting from the interaction of UIM-SH3 and AMSH-P2 is relatively narrow."
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"We can thus rule out the fact that two AMSH-P2 could bind SH3."
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"To a Lys63-Ub 2 sample, which was 15 N-labeled on the distal domain and saturated by UIM-SH3, we added AMSH-P2 until saturation."
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"We performed similar experiments for the proximal domain of a 15 N-Lys63-Ub 2 sample saturated first by UIM-SH3 and then by AMSH-P2."
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"First and foremost, we were seeking to determine if AMSH-P2 shows a well-defined secondary structure."
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"The use of the PrDos server predicted the AMSH-P2 sequence as unstructured (see Fig. S20)."
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"From a series of 1D 1 H, 2D 1 H, 1 H total correlated spectroscopy (TOCSY), and 2D 1 H, 1 H nuclear Overhauser enhancement (NOE) spectroscopy (NOESY) experiments, we assigned the AMSH-P2 backbone and [MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"To determine whether direct contacts exist between AMSH-P2 and the SH3 domain of STAM2, we acquired 3D 15 N-filtered, 15 N-edited NOESY, 3D 13 C-filtered, and 13 C-edited NOESY experiments on a 13 C, [MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"Although AMSH-P2 is unstructured in the free state, intermolecular NOEs resulting from the above experiments were observed between the backbone or side-chain protons of the SH3 domain and the side-cha[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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"To allow an accurate assignment of NOEs, we reassigned the backbone and side-chain proton resonances of UIM-SH3 in complex with AMSH-P2 by a series of 3D experiments (see Material and Methods )."
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"The HN, N, CO, Cα, Cβ, and Hα resonance assignments of UIM-SH3 bound to AMSH-P2 were obtained and subjected to Chemical Shift Index (CSI) analysis [38] ."
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"The resulting analysis showed that UIM-SH3 did not experience any critical conformational rearrangement upon binding to AMSH-P2 and keeps a similar fold as in the free state (Figs."
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"The AMSH-P2 proton resonance assignments were also determined by filtering out the protein NMR signals (see Material and Methods)."
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"Importantly, the strongest intermolecular NOEs were observed between the Lys231, Tyr218, and Trp246 residues of SH3 and the Val233, Arg235, and Pro239 residues of AMSH-P2."
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"Figure 4 b shows that AMSH-P2 is wedged in a groove delimited by the RT loop, the n-Src loop, and the 3 10 helix of SH3."
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"The AMSH-P2 peptide assignment in the free state was carried out by using a combination of the following experiments: { 1 H, 1 H}-TOCSY and { 1 H, 1 H}-NOESY experiments."
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"Assignment and intramolecular NOEs of the unlabeled AMSH-P2 in complex with UIM-SH3 were obtained from 2D TOCSY and NOESY ( 15 N, 13 C)-filtered (mixing time equal to 60 ms for TOCSY and 120 ms for NO[MISSING/INVALID CREDENTIALS: limited to 200 char for Elsevier]"
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