Calmodulin (CaM) binds most of its targets by wrappingaround an amphipathic ?-helix. The N-terminus of Orai proteins contains a conserved CaM-binding segment but the binding mechanism is only partially characterized. Here, microscale thermophoresis (MST), surface plasmon resonance (SPR), andatomic force microscopy (AFM) were employed to study the binding equilibria, the kinetics, and the single-molecular interaction forces involved in the binding of CaM to the conserved helical segments of Orai1 and Orai3. The results consistently indicated step-wise binding of two separate target peptides to the two lobes of CaM. An unparalleled high affinity was found when two Orai peptides were dimerized or immobilized at high lateral density, thereby mimicking the close proximity of the N-termini in native Orai oligomers. The analogous experiments with smooth muscle myosin light chain kinase (smMLCK) showed only the expected 1:1 binding, confirming the validity of our methods.
[NMR paper] Use of Fluorine Pseudocontact Shifts to Characterize the Protein-Ligand Interaction Mode in the Limit of NMR Intermediate Exchange.
Use of Fluorine Pseudocontact Shifts to Characterize the Protein-Ligand Interaction Mode in the Limit of NMR Intermediate Exchange.
Use of Fluorine Pseudocontact Shifts to Characterize the Protein-Ligand Interaction Mode in the Limit of NMR Intermediate Exchange.
Angew Chem Int Ed Engl. 2017 Aug 28;:
Authors: Gao J, Liang E, Ma R, Li F, Liu Y, Liu J, Jiang L, Li C, Dai H, Wu J, Su X, He W, Ruan K
Abstract
The characterization of protein-ligand interaction modes becomes recalcitrant in the NMR intermediate exchange regime as the...
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[NMR paper] Use of Fluorine Pseudocontact Shifts to Characterize the Protein-Ligand Interaction Mode in the Limit of NMR Intermediate Exchange
Use of Fluorine Pseudocontact Shifts to Characterize the Protein-Ligand Interaction Mode in the Limit of NMR Intermediate Exchange
The characterization of protein-ligand interaction modes becomes recalcitrant in the NMR intermediate exchange regime as the interface resonances are broadened beyond detection. Here, we determined the 19F low-populated bound-state pseudocontact shifts (PCSs) of mono- and di-fluorinated inhibitors of the BRM bromodomain using a highly-skewed protein/ligand ratio. The bound-state 19F PCSs were retrieved from 19F chemical exchange saturation transfer (CEST) in...
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Mode of Interaction of the Signal-Transducing ProteinEIIAGlc with the Maltose ABC Transporter in the Processof Inducer Exclusion
Mode of Interaction of the Signal-Transducing ProteinEIIAGlc with the Maltose ABC Transporter in the Processof Inducer Exclusion
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/bichaw/0/bichaw.ahead-of-print/acs.biochem.6b00721/20160919/images/medium/bi-2016-007219_0007.gif
Biochemistry
DOI: 10.1021/acs.biochem.6b00721
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Free-electron lasers reveal detailed architecture of proteins - R & D Magazine
Free-electron lasers reveal detailed architecture of proteins - R & D Magazine
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R & D Magazine
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Free-electron lasers reveal detailed architecture of proteins
R & D Magazine
An international team of researchers has recently analyzed protein crystals using ... of a new 0.75-mm solid state nuclear magnetic resonance (NMR) probe.
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NMR structural study of the intracellular loop 3 of the serotonin 5-HT(1A) receptor and its interaction with calmodulin.
NMR structural study of the intracellular loop 3 of the serotonin 5-HT(1A) receptor and its interaction with calmodulin.
NMR structural study of the intracellular loop 3 of the serotonin 5-HT(1A) receptor and its interaction with calmodulin.
Biochim Biophys Acta. 2011 May 24;
Authors: Chen AS, Kim YM, Gayen S, Huang Q, Raida M, Kang C
The serotonin (5-HT(1A)) receptor, a G-protein-coupled receptor (GPCR), plays important roles in serotonergic signaling in the central nervous system. The third intracellular loop (ICL3) of the 5-HT(1A) receptor has...
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[NMR paper] NMR and molecular dynamics studies of the interaction of melatonin with calmodulin.
NMR and molecular dynamics studies of the interaction of melatonin with calmodulin.
Related Articles NMR and molecular dynamics studies of the interaction of melatonin with calmodulin.
Protein Sci. 2004 Nov;13(11):2925-38
Authors: Turjanski AG, Estrin DA, Rosenstein RE, McCormick JE, Martin SR, Pastore A, Biekofsky RR, Martorana V
Pineal hormone melatonin (N-acetyl-5-methoxytryptamine) is thought to modulate the calcium/calmodulin signaling pathway either by changing intracellular Ca(2+) concentration via activation of its G-protein-coupled...
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11-24-2010 10:03 PM
[NMR paper] 13C NMR study of the mode of interaction in solution of the B fragment of staphylococ
13C NMR study of the mode of interaction in solution of the B fragment of staphylococcal protein A and the Fc fragments of mouse immunoglobulin G.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles 13C NMR study of the mode of interaction in solution of the B fragment of staphylococcal protein A and the Fc fragments of mouse immunoglobulin G.
FEBS Lett. 1993 Aug 9;328(1-2):49-54
Authors: Kato K, Gouda H, Takaha W, Yoshino A, Matsunaga C, Arata Y
The mode of interaction of...
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[NMR paper] Interaction of calmodulin with phospholamban and caldesmon: comparative studies by 1H
Interaction of calmodulin with phospholamban and caldesmon: comparative studies by 1H-NMR spectroscopy.
Related Articles Interaction of calmodulin with phospholamban and caldesmon: comparative studies by 1H-NMR spectroscopy.
Biochim Biophys Acta. 1992 Nov 10;1160(1):22-34
Authors: Gao Y, Levine BA, Mornet D, Slatter DA, Strasburg GM
In order to identify comparative aspects of the interaction of calmodulin with its target proteins, proton magnetic-resonance studies of complex formation between calmodulin and defined segments of phospholamban...
Detailed Evidence for an Unparalleled Interaction Mode between Calmodulin and Orai Proteins
Linear Mode
