Human multiprotein bridging factor 1 and Calmodulin do not interact in vitro as confirmed by NMR spectroscopy and CaM-agarose affinity chromatography.
Protein Expr Purif. 2011 Jul 14;
Authors: Babini E, Hu X, Parigi G, Vignali M
The human multiprotein bridging factor 1 (hMBF1) has been established in different cellular types to have the role of transcriptional coactivator. It is also reported to be a putative Calmodulin (CaM) target, able to bind CaM in its calcium-free state, but little is known about the structural features and the biological relevance of this interaction. We applied NMR to investigate the interaction between the two proteins in solution and compared the results with those obtained with CaM-agarose affinity chromatography. No changes in (1)H-(15)N HSQC spectrum of both apo-CaM and Ca(2+)-CaM upon addition of hMBF1 prove that the two proteins do not interact in vitro. These results were confirmed by CaM-agarose affinity chromatography when operating under the same conditions. The discrepancy between present and previous experiments performed with CaM-agarose affinity chromatography depends on different experimental parameters suggesting that particular attention must be paid when CaM, or other immobilized proteins, are used to measure their affinity with putative partners. These results also imply that if an interaction between the two proteins exists in vivo, as reported for hMBF1 of endothelial cells, it might involve a posttranslational modified form of the proteins or it relies on other conditions imposed by the cellular environment.
PMID: 21782027 [PubMed - as supplied by publisher]
Solution NMR Structure of apo-calmodulin in complex with the IQ motif of Human Cardiac Sodium Channel Na(V)1.5.
Solution NMR Structure of apo-calmodulin in complex with the IQ motif of Human Cardiac Sodium Channel Na(V)1.5.
Solution NMR Structure of apo-calmodulin in complex with the IQ motif of Human Cardiac Sodium Channel Na(V)1.5.
J Mol Biol. 2010 Dec 14;
Authors: Chagot B, Chazin WJ
The function of the human voltage-gated sodium channel Na(V)1.5 is regulated in part by intracellular calcium signals. The ubiquitous calcium sensor protein calmodulin (CaM) is an important part of the complex calcium-sensing apparatus in Na(V)1.5. CaM interacts with an IQ...
[NMR paper] NMR studies of a murine-human chimera of leukaemia inhibitory factor (LIF). Compariso
NMR studies of a murine-human chimera of leukaemia inhibitory factor (LIF). Comparison with human LIF.
Related Articles NMR studies of a murine-human chimera of leukaemia inhibitory factor (LIF). Comparison with human LIF.
Growth Factors. 1994;11(4):271-6
Authors: Maurer T, Smith DK, Owczarek CM, Layton MJ, Zhang JG, Nicola NA, Norton RS
Leukaemia inhibitory factor (LIF) is a polyfunctional cytokine active on many cell types. We present here 1H NMR studies on the solution properties and stability of MH35, a chimera of murine and human LIF...
nmrlearner
Journal club
0
08-22-2010 03:33 AM
[NMR paper] Structure and dynamics of the human granulocyte colony-stimulating factor determined
Structure and dynamics of the human granulocyte colony-stimulating factor determined by NMR spectroscopy. Loop mobility in a four-helix-bundle protein.
Related Articles Structure and dynamics of the human granulocyte colony-stimulating factor determined by NMR spectroscopy. Loop mobility in a four-helix-bundle protein.
Biochemistry. 1994 Jul 19;33(28):8453-63
Authors: Zink T, Ross A, Lüers K, Cieslar C, Rudolph R, Holak TA
Recombinant 15N- and 13C-labeled human granulocyte colony-stimulating factor (rh-metG-CSF) has been studied by 2D and 3D...
nmrlearner
Journal club
0
08-22-2010 03:29 AM
[NMR paper] Comparative studies of the interaction of human and bovine platelet factor 4 with hep
Comparative studies of the interaction of human and bovine platelet factor 4 with heparin using histidine NMR resonances as spectroscopic probes.
Related Articles Comparative studies of the interaction of human and bovine platelet factor 4 with heparin using histidine NMR resonances as spectroscopic probes.
J Protein Chem. 1993 Jun;12(3):303-9
Authors: Talpas CJ, Lee L
The pKa values of His-38 and His-50 of the heparin-binding protein, bovine platelet factor 4, are 5.6 and 6.5, respectively, as determined by 1H NMR spectroscopy. The 1H NMR...
nmrlearner
Journal club
0
08-21-2010 11:53 PM
[NMR paper] 1H-NMR assignment and secondary structure of human insulin-like growth factor-I (IGF-
1H-NMR assignment and secondary structure of human insulin-like growth factor-I (IGF-I) in solution.
Related Articles 1H-NMR assignment and secondary structure of human insulin-like growth factor-I (IGF-I) in solution.
J Biochem. 1992 Apr;111(4):529-36
Authors: Sato A, Nishimura S, Ohkubo T, Kyogoku Y, Koyama S, Kobayashi M, Yasuda T, Kobayashi Y
Human insulin-like growth factor-I (IGF-I) was studied by two-dimensional 1H-NMR spectroscopy. Resonance assignments were obtained for all the backbone protons and almost all of the sidechain protons...
nmrlearner
Journal club
0
08-21-2010 11:41 PM
[NMR paper] Low-affinity platelet factor 4 1H NMR derived aggregate equilibria indicate a physiol
Low-affinity platelet factor 4 1H NMR derived aggregate equilibria indicate a physiologic preference for monomers over dimers and tetramers.
Related Articles Low-affinity platelet factor 4 1H NMR derived aggregate equilibria indicate a physiologic preference for monomers over dimers and tetramers.
Biochemistry. 1991 Jan 29;30(4):925-34
Authors: Mayo KH
Low-affinity platelet factor 4 (LA-PF4), unlike another related, sequentially homologous (about 50%) platelet-specific protein, platelet factor 4 (PF4), is an active mitogenic and chemotactic...
nmrlearner
Journal club
0
08-21-2010 11:16 PM
[NMR paper] Solution structures of human transforming growth factor alpha derived from 1H NMR dat
Solution structures of human transforming growth factor alpha derived from 1H NMR data.
Related Articles Solution structures of human transforming growth factor alpha derived from 1H NMR data.
Biochemistry. 1990 Aug 28;29(34):7805-13
Authors: Kline TP, Brown FK, Brown SC, Jeffs PW, Kopple KD, Mueller L
The 600-MHz 1H NMR spectrum of the des-Val-Val mutant of human transforming growth factor alpha (TGF-alpha) was reassigned at pH = 6.3. The conformation space of des-Val-Val TGF-alpha was explored by distance geometry embedding followed by...
Human multiprotein bridging factor 1 and Calmodulin do not interact in vitro as confirmed by NMR spectroscopy and CaM-agarose affinity chromatography.
Linear Mode
