Na<sup>+</sup> binding to parvalbumins studied by <sup>23</sup>Na NMR

Parello, Joseph; Reimarsson, Pétur; Thulin, Eva; Lindman, Björn

doi:10.1016/0014-5793(79)81153-9

Cited by 23 publications

(9 citation statements)

References 17 publications

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“…It must be noted though that in this and other studies on parvalbumins, which find evidence for Mg*+ binding to CD and EF sites, soluble chelators such as EGTA or EDTA have been used to remove Ca*' from the native protein. It has been shown that EGTA is able to bind to apoparvalbumin and thereby to interact strongly with different cations such as Ca*+ and Na' [22] . In view of this we make presently noattempt to correlate our data, obtained without using a soluble chelator, with literature data on cation binding to parvalbumins.…”

Section: Discussionmentioning

confidence: 99%

Mg²⁺ binding to parvalbumins studied by ²⁵Mg and ¹¹³Cd NMR spectroscopy

Cavé

Parello

Drakenberg³

et al. 1979

FEBS Letters

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Section: Discussionmentioning

confidence: 99%

Mg²⁺ binding to parvalbumins studied by ²⁵Mg and ¹¹³Cd NMR spectroscopy

Cavé

Parello

Drakenberg³

et al. 1979

FEBS Letters

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“…By contrast, Parello et al (1979) observed only a very slight 23Na excess linewidth in the presence of hake parvalbumin, where the Ca2+ ions had been fully or partly removed. It was observed, however, that the addition of the complexing agent EGTA to the parvalbumin solutions caused a marked increase of the 23Na excess linewidth and that this effect could be reversed by the addition of CaZ+.…”

Section: B Survey Of Experimental Workmentioning

confidence: 77%

“…Under the assumption that x = 0.5 MHz (see the discussion of sodium binding to polyelectrolytes in Section 111.4), one may calculate that 20 Na+ ions are bonded per oxygen carrying hemocyanin subunit in the absence of Ca2+ (Norne et al, 1979a). Sodium ion binding to parvalbumins has been studied in two laboratories (Grandjean et al, 1977;Parello et al, 1979). Parvalbumins are a class of low molecular weight proteins (about 11,500 daltons) found in the muscles of most vertebrates.…”

Section: B Survey Of Experimental Workmentioning

confidence: 99%

“…EGTA is commonly employed in the preparation of calcium-free parvalbumins. Parello et al (1979) deliberately avoided the use of this complexing agent and it would appear possible that some EGTA may inadvertently have been present in the preparations used by Grandjean et al (1977. 3sK+ has been virtually unexplored to study K+-protein interactions with the exception of an exploratory study by , who observed enhanced 3sK+ relaxation rates in the presence of pyruvate kinase.…”

Section: B Survey Of Experimental Workmentioning

confidence: 99%

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Ion Binding in Biological Systems as Studied by NMR Spectroscopy

Forsén

Lindman

1981

Methods of Biochemical Analysis

101

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“…In order to avoid any interference from a strong chelator (such as EDTA) in the NMR cation-binding studies [23] the bone was demineralized in acetic acid instead of using the more common EDTA procedure. The isolated protein was characterized by SDS polyacrylamide gel electrophoresis, HPLC, amino acid analysis and sequence determination.…”

Section: Preparation and Characterization Of Bgpmentioning

confidence: 99%

Calcium binding to bone gamma-carboxyglutamic acid protein from calf studied by 43Ca NMR

Drakenberg

Andersson

et al. 1986

Eur J Biochem

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Calcium binding to bone y-carboxyglutamic acid protein (BGB) from calf has been studied using 43Ca NMR. The temperature dependence of the 43Ca NMR signal has been used to calculate the calcium ion exchange rate, koff. The dependence of the 43Ca NMR band shape on the [Ca2+]/[BGP] ratio fits well to a chemical equilibrium model having a single Ca2+-binding site with an association constant in the range of 5 x lo3 -1 x lo5 M-I. The pH dependence of the 43Ca NMR line-width shows a single apparent pK, value of 5.1.Bone from mammals, birds and fishes contains a lowmolecular-mass non-collagenous protein, in which the unusual amino acid y-carboxyglutamic acid is found [l, 21. The protein, which has been named osteocalcin or bone y-carboxyglutamic acid protein (BGP), consists of a single polypeptide chain of about 50 amino acid residues, two or three of which are y-carboxyglutamic acid [3 -51. y-Carboxyglutamic acid, which is also found in some of the coagulation factors and related plasma proteins [6], is formed by a vitamin-K-dependent posttranslational carboxylation of certain glutamic acid residues in the protein polypeptide [7, 81. Most of the BGP is located in the extracellular bone matrix, adsorbed to the mineral crystals [9-121, but BGP also occurs in plasma at low concentrations [13- Abbreviations. Gla, 9-carboxyglutamic acid; BGP, bone Gla protein, osteocalcin; HPLC, high-performance liquid chromatography; SDS, sodium dodecyl sulphate.Note. The rotational correlation time ( 7 : ' ) of BGP can be estimated to approximately 2 ns at 20°C from the Debye-StokesEinstein equation [25]. We are thus able to determine an upper limit of 02, in our experiments: w t 2 <0.22 (w = 1.078. 10' s-' and ~~ rc <2 . 10-9 s~.technique, which has proven to be very useful in the study of various kinds of calcium-binding protein. BGP from calf bone was chosen because it is well characterized [3] and calf bone can easily be obtained in large quantities. A simple procedure for the large-scale purification of BGP from calf bone is also presented. MATERIALS AND METHODS Preparation and characterization of BGPBGP was isolated from calf bone by a large-scale purification procedure. In order to avoid any interference from a strong chelator (such as EDTA) in the NMR cation-binding studies [23] the bone was demineralized in acetic acid instead of using the more common EDTA procedure. The isolated protein was characterized by SDS polyacrylamide gel electrophoresis, HPLC, amino acid analysis and sequence determination. The isolated BGP was judged to be at least 95% pure and the analytical data, including Gla content, were in good agreement with published data on the protein [2,3]. The details of the purification procedure and the results of the characterization of the purified protein are given in the miniprint supplement. BGP solutions for NMR measurementsCa2 + -free BGP was prepared by passing an aqueous solution of the protein through a column of Chelex-100 (BioRad Labs., Richmond, CA, USA) resulting in a protein containing less than 0.1 equ...

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Na⁺ binding to parvalbumins studied by ²³Na NMR

Cited by 23 publications

References 17 publications

Mg²⁺ binding to parvalbumins studied by ²⁵Mg and ¹¹³Cd NMR spectroscopy

Mg²⁺ binding to parvalbumins studied by ²⁵Mg and ¹¹³Cd NMR spectroscopy

Ion Binding in Biological Systems as Studied by NMR Spectroscopy

Calcium binding to bone gamma-carboxyglutamic acid protein from calf studied by 43Ca NMR

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Na+ binding to parvalbumins studied by 23Na NMR

Cited by 23 publications

References 17 publications

Mg2+ binding to parvalbumins studied by 25Mg and 113Cd NMR spectroscopy

Mg2+ binding to parvalbumins studied by 25Mg and 113Cd NMR spectroscopy

Ion Binding in Biological Systems as Studied by NMR Spectroscopy

Calcium binding to bone gamma-carboxyglutamic acid protein from calf studied by 43Ca NMR

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Na⁺ binding to parvalbumins studied by ²³Na NMR

Mg²⁺ binding to parvalbumins studied by ²⁵Mg and ¹¹³Cd NMR spectroscopy

Mg²⁺ binding to parvalbumins studied by ²⁵Mg and ¹¹³Cd NMR spectroscopy