Protein L is a cell wall protein expressed by some strains of the anaerobic bacterial species Peptostreptococcus magnus. It binds to immunoglobulin (Ig) light chains predominantly of the kappa subtype from a wide range of animal species. This binding is mediated by five highly homologous repeats designated as B1-B5, each of which comprises 72 to 76 amino acid residues. The fold of the Ig light chain-binding B1 domain of protein L has previously been shown to comprise an alpha-helix packed against a four-stranded beta-sheet. The Ig-binding region of the protein L domain involves most of the residues in the second beta-strand, the C-terminal residues of the alpha-helix, and residues in the loop connecting the alpha-helix with the third beta-strand. In the present study, we have identified the protein L-binding site of an Ig light chain by use of stable isotope-assisted NMR spectroscopy. The light chain of a murine monoclonal anti-17alpha-hydroxy-progesterone Fab fragment (IgG2b, kappa) was selectively labeled with 13C at carbonyl groups of Ala, Arg, Cys, Ile, Lys, Met, Phe, Trp, or Tyr. The residues in which the carbonyl 13C chemical shift was significantly perturbed upon binding of the protein L B1 domain were preferentially found in the second beta-strand of the variable kappa domain and parts of its flanking beta-strands. None of these residues were affected by the addition of the antigen against which the monoclonal Fab fragment is directed. Therefore, we conclude that protein L binds to the outer surface of the framework region of the V(L) domain, primarily involving the V(L) second strand, and that this binding is independent of antigen-binding. The present NMR data, in combination with sequence comparisons between kappa light chains with and without protein L affinity, suggest that the amino acid substitutions at positions 9, 20, and/or 74 of the kappa light chains could crucially affect the interaction between protein L and the V(L) domain.
Solid-state NMR analysis of interaction sites of curcumin and 42-residue amyloid ?-protein fibrils.
Solid-state NMR analysis of interaction sites of curcumin and 42-residue amyloid ?-protein fibrils.
Solid-state NMR analysis of interaction sites of curcumin and 42-residue amyloid ?-protein fibrils.
Bioorg Med Chem. 2011 Aug 27;
Authors: Masuda Y, Fukuchi M, Yatagawa T, Tada M, Takeda K, Irie K, Akagi KI, Monobe Y, Imazawa T, Takegoshi K
Abstract
Aggregation of 42-residue amyloid ?-protein (A?42) plays a pivotal role in the etiology of Alzheimer's disease (AD). Curcumin, the yellow pigment in the rhizome of turmeric, attracts...
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First solid-state NMR analysis of uniformly (13)C-enriched major light-harvesting complexes from Chlamydomonas reinhardtti and identification of protein and cofactor spin clusters.
First solid-state NMR analysis of uniformly (13)C-enriched major light-harvesting complexes from Chlamydomonas reinhardtti and identification of protein and cofactor spin clusters.
First solid-state NMR analysis of uniformly (13)C-enriched major light-harvesting complexes from Chlamydomonas reinhardtti and identification of protein and cofactor spin clusters.
Biochim Biophys Acta. 2011 Jan 25;
Authors: Pandit A, Morosinotto T, Reus M, Holzwarth AR, Bassi R, de Groot HJ
The light-harvesting complex II (LHCII) is the main component of the...
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[NMR paper] Biosynthetic site-specific (13) C labeling of the light-harvesting 2 protein complex:
Biosynthetic site-specific (13) C labeling of the light-harvesting 2 protein complex: a model for solid state NMR structure determination of transmembrane proteins.
Related Articles Biosynthetic site-specific (13) C labeling of the light-harvesting 2 protein complex: a model for solid state NMR structure determination of transmembrane proteins.
J Biomol NMR. 2004 Nov;30(3):267-74
Authors: van Gammeren AJ, Hulsbergen FB, Hollander JG, de Groot HJ
Partly biosynthetic site-directed isotopically (13)C enriched photosynthetic light-harvesting...
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11-24-2010 10:03 PM
[NMR paper] The effects of mutations on motions of side-chains in protein L studied by 2H NMR dyn
The effects of mutations on motions of side-chains in protein L studied by 2H NMR dynamics and scalar couplings.
Related Articles The effects of mutations on motions of side-chains in protein L studied by 2H NMR dynamics and scalar couplings.
J Mol Biol. 2003 Jun 6;329(3):551-63
Authors: Millet O, Mittermaier A, Baker D, Kay LE
Recently developed 2H spin relaxation experiments are applied to study the dynamics of methyl-containing side-chains in the B1 domain of protein L and in a pair of point mutants of the domain, F22L and A20V. X-ray and...
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[NMR paper] Analysis of protein-carbohydrate interaction at the lower size limit of the protein p
Analysis of protein-carbohydrate interaction at the lower size limit of the protein part (15-mer peptide) by NMR spectroscopy, electrospray ionization mass spectrometry, and molecular modeling.
Related Articles Analysis of protein-carbohydrate interaction at the lower size limit of the protein part (15-mer peptide) by NMR spectroscopy, electrospray ionization mass spectrometry, and molecular modeling.
Biochemistry. 2002 Jul 30;41(30):9707-17
Authors: Siebert HC, Lü SY, Frank M, Kramer J, Wechselberger R, Joosten J, André S, Rittenhouse-Olson K, Roy R, von...
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[NMR paper] Side-chains in native and random coil protein conformations. Analysis of NMR coupling
Side-chains in native and random coil protein conformations. Analysis of NMR coupling constants and chi1 torsion angle preferences.
Related Articles Side-chains in native and random coil protein conformations. Analysis of NMR coupling constants and chi1 torsion angle preferences.
J Mol Biol. 1998 Jul 31;280(5):867-77
Authors: West NJ, Smith LJ
The behaviour of amino acid side-chains in proteins in solution has been characterised by analysing NMR 3JHalphaH beta coupling constants and crystallographic chi1 torsion angles. Side-chains both in the...
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[NMR paper] Assignment of protein NMR spectra in the light of homonuclear 3D spectroscopy: an aut
Assignment of protein NMR spectra in the light of homonuclear 3D spectroscopy: an automatable procedure based on 3D TOCSY-TOCSY and 3D TOCSY-NOESY.
Related Articles Assignment of protein NMR spectra in the light of homonuclear 3D spectroscopy: an automatable procedure based on 3D TOCSY-TOCSY and 3D TOCSY-NOESY.
Biopolymers. 1991 May;31(6):699-712
Authors: Oschkinat H, Holak TA, Cieslar C
Automated assignment of proteins is greatly simplified using data from 3D-nmr spectra. A strategy is presented which makes use of 3D-TOCSY-TOCSY and...
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08-21-2010 11:16 PM
[NMR paper] NMR analysis of the structure of synaptobrevin and of its interaction with syntaxin.
NMR analysis of the structure of synaptobrevin and of its interaction with syntaxin.
Related Articles NMR analysis of the structure of synaptobrevin and of its interaction with syntaxin.
J Biomol NMR. 1999 Jul;14(3):203-7
Authors: Hazzard J, Südhof TC, Rizo J
Synaptobrevin is a synaptic vesicle protein that has an essential role in exocytosis and forms the SNARE complex with syntaxin and SNAP-25. We have analyzed the structure of isolated synaptobrevin and its binary interaction with syntaxin using NMR spectroscopy. Our results demonstrate...