Changes in the 4-PDS-reactive —SH groups of hemoglobin associated with the binding of phosphates and ligands

Taketa, F.; Morell, Samuel A.

doi:10.1016/0003-2697(69)90119-5

Cited by 18 publications

(13 citation statements)

References 8 publications

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“…Two reactive thiol groups/ tetramer are observed in both the cross-linked and the native forms of HbA 0 . As previously observed (Taketa and Morell, 1969) and confirmed here for native hemoglobin, conversion of the cross-linked hemoglobins to the carbonyl derivative slightly decreases the reactivity with DTDP relative to that of the corresponding oxyhemoglobin derivatives. Furthermore, deoxygenation of the cross-linked hemoglobins decreased reactivity of the reactive thiol as previously observed for the native protein (Riggs, 1961;Morell et al, 1962;Benesch and Benesch, 1962).…”

Section: Sulfhydryl Reactivitysupporting

confidence: 89%

“…The reactivity of the ␤93Cys residue in native and cross-linked forms of hemoglobin was studied by monitoring the reaction of these proteins with 4,4Ј-dithiodipyridine (DTDP) (Aldrich Chemical Co., Milwaukee, WI) (Taketa and Morell, 1969). In these measurements, the protein solution (2.5 ml, 10 M in heme) in bis/Tris buffer (50 mM, pH 7.0) or N-tris(hydroxyethyl)methyl-3-aminopropane sulfonic acid buffer (50 mM, pH 8.4) was mixed with 0.2 ml of DTDP solution (12.5 mM in the same buffer as the protein).…”

Section: Sulfhydryl Reactivitymentioning

confidence: 99%

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Functional Comparison of Specifically Cross-Linked Hemoglobins Biased Toward the R and T States

Johnson

Adamson

Mauk

1998

Biophysical Journal

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Selected functional and spectroscopic properties of two human hemoglobin (HbA0) derivatives that were site-specifically cross-linked in the cleft between beta-chains where 2, 3-bisphosphoglycerate normally binds have been determined to assess the effects of the cross-linking on the behavior of the protein. Trimesoyl tris(3,5-dibromosalicylate) (TTDS) cross-links Hb between beta82Lys residues. The resulting TTDS-Hb exhibits a slower rate of oxygen dissociation and an increased rate of carbon monoxide association than observed for HbA0. The electron paramagnetic resonance (EPR) spectrum of TTDS-HbNO does not exhibit the hyperfine structure that is indicative of significant conformational change despite the fact that the 2,3-bisphosphoglycerate binding site is occupied by the cross-linking reagent. The reactivity of the beta93Cys residues of TTDS-Hb is only slightly decreased relative to that of HbA0. On the other hand, cross-linking Hb between Lys82 and the amino-terminal beta1Val group with trimesoyl tris(methyl phosphate) (TMMP) increases the rate of oxygen dissociation and reduces the rate of CO association relative to the rates observed for HbA0. In addition, the EPR spectrum of the TMMP-HbNO exhibits the three-line hyperfine structure that results from disruption of the proximal His-Fe bond of the alpha-chains, and the accessibility of the betaCys93 residues in this derivative is decreased fourfold. The present results are consistent with the conclusion that the quaternary structure of TTDS-Hb is shifted toward the R state whereas the quaternary structure of TMMP-Hb is shifted toward the T state and provides additional evidence that the identity of the residues involved in intramolecular cross-linking of hemoglobin within the 2,3-bisphosphoglycerate binding site between beta-chains can have a significant influence on the conformational and functional properties of the protein.

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Section: Sulfhydryl Reactivitysupporting

confidence: 89%

Section: Sulfhydryl Reactivitymentioning

confidence: 99%

Functional Comparison of Specifically Cross-Linked Hemoglobins Biased Toward the R and T States

Johnson

Adamson

Mauk

1998

Biophysical Journal

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“…Hemoglobin Solutions. Hemolysates were prepared from washed erythrocytes as previously described (Taketa and Morell, 1969). Hb Wood was isolated by CM-Sephadex column chromatography using 0.05 M phosphate buffer (pH 6.7) at 4°( F. Taketa et al, submitted for publication).…”

Section: Methodsmentioning

confidence: 99%

“…Reaction was initiated by tipping 0.2 ml of the PDS solution (0.44 mg/ml) from the side arm into the cuvet compartment containing 2.0 ml of 0.1% Hb solution (in 0.05 M bis-tris, 0.1 M NaCl, and 0.1 mM EDTA (pH 7.0)) and 0.3 ml of the buffer with or without IHP. The reaction was followed at 324 nm with a Gilford Model 240 recording spectrophotometer as previously described (Taketa and Morell, 1969).…”

Section: Methodsmentioning

confidence: 99%

Nitrosylhemoglobin wood. Effects of inositol hexaphosphate on thiol reactivity and electron paramagnetic resonance spectrum

Taketa¹,

Antholine²,

Mauk³

et al. 1975

Biochemistry

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Properties of Hb Wood (beta-97(FG4)His leads to Leu), a high oxygen affinity hemoglobin with reduced hemeheme interaction, were examined in its nitric oxide liganded form. The reactivity of the beta-93 thiol groups and the electron paramagnetic resonance (EPR) spectrum were examined to determine what effect the amino acid substitution, which occurs at the alpha1beta2 interface, would have on inositol hexaphosphate induced transition of this form of the tetramer. Binding of inositol hexaphosphate (IHP) in a 1:1 stoichiometry was demonstrated. In spite of apparently normal interaction with IHP, there was little or no change in the reactivity of the beta-93 thiol groups and in the electron paramagnetic resonance (EPR) spectrum as contrasted with the marked changes characteristic of normal hemoglobin (HbA). In contrast with NO-HbA, there was also no development of the EPR hyperfine structure in NO-Hb Wood with increased protonation of the protein at pH below 7.0. Taken together with the observations of Henry and Banerjee ((1973), J. Mol. Biol. 73, 469) on the development of NO-Hb EPR hyperfine structure and of Perutz et al. (1974a), Biochemistry 13, 2174) on changes in thiol reactivity with the R leads to T transition, the results suggest that IHP or H+ cannot switch NO-Hb Wood to the T conformation. Since the atomic structures of met- and deoxyhemoglobin offer no indication that His-97 plays any special part in the allosteric mechanism (M. E. Perutz, personal communication), it appears that the replacement of His-97 by Leu reduces the stability of the T structure relative to that of R.

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“…Alternatively, zinc might bind to only one of these residues and the observed effects involving the other residue would be mediated through conformational changes linked to the binding of zinc. Consistent with this possibility is the effect of 2,3-DPG on the reaction of 4,4'-dipyridine disulfide (4PDS) with cysteine-093 (Taketa and Morell, 1969). 2,3-DPG binds to hemoglobin at the entrance to the central cavity between the two 0 chains (Amone, 1972).…”

Section: Resultsmentioning

confidence: 97%

Interaction of zinc with hemoglobin: binding of zinc and the oxygen affinity

Rifkind¹,

Heim²

1977

Biochemistry

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Stripped human hemoglobin was shown to have a high apparent zinc association constant of 1.3 X 10(7) M-1 with a stoichiometry of one zinc for every two hemes. The saturation of this site produces a dramatic 3.7-fold increase in the oxygen affinity. The effect of zinc on the oxygen affinity is interrelated with the interaction of 2,3-diphosphoglyceric acid (2,3-DPG) and hemoglobin. Thus, a smaller zinc effect is observed in the presence of added 2,3-DPG. Information about the location of the zinc-binding site responsible for the increased oxygen affinity has been obtained by comparing the binding of zinc to various hemoglobins. Blocking the beta93 sulfhydryl group decreases the apparent zinc association constant by an order of magnitude. The substitution of histidine-beta143 in hemoglobin Abruzzo [beta143 (H21) His leads to Arg] and hemoglobin Little Rock [beta143 (H21) His leads to Gln] decreases the apparent zinc association constant by two orders of magnitude. The substitution of histidine-beta143 by other amino acids and the reaction of the beta93 sulfhydryl group are known to produce dramatic increases in the oxygen affinity. The binding of zinc to one or both of these amino acids can, therefore, explain the zinc-induced increase in the oxygen affinity.

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Changes in the 4-PDS-reactive —SH groups of hemoglobin associated with the binding of phosphates and ligands

Cited by 18 publications

References 8 publications

Functional Comparison of Specifically Cross-Linked Hemoglobins Biased Toward the R and T States

Functional Comparison of Specifically Cross-Linked Hemoglobins Biased Toward the R and T States

Nitrosylhemoglobin wood. Effects of inositol hexaphosphate on thiol reactivity and electron paramagnetic resonance spectrum

Interaction of zinc with hemoglobin: binding of zinc and the oxygen affinity

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