Positive and negative cooperativities at subsequent steps of oxygenation regulate the allosteric behavior of multistate sebacylhemoglobin

Enrico Bucci, Anna Razynska, Herman Kwansa, Zygmunt Gryczynski, John H. Collins, Clara Fronticelli, Ron Unger, Michael Braxenthaler, John Moult, Xinhua Ji, Gary Gilliland

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Abstract

Cross-linked human hemoglobin (HbA) is obtained by reaction with bis(3,5- dibromosalicyl) sebacate. Peptide maps and crystallographic analyses confirm the presence of the 10 carbon atom long sebacyl residue cross-linking the two β82 lysines of the β-cleft (DecHb). The Adair's constants, obtained from the oxygen binding isotherms, show that at the first step of oxygenation normal hemoglobin and DecHb have a very similar oxygen affinity. In DecHb negative binding cooperativity is present at the second step of oxygenation, which has an affinity 27 times lower than at the first step. Positive cooperativity is present at the third binding step, whose affinity is 380 times that of the second step. The fourth binding step shows a weak negative cooperativity with an affinity one-half that of the third step. Crystals of deoxy-DecHb diffracted to 1.9 Å resolution. The resulting atomic coordinates are very similar to those of Fermi et al. [(1984) J. Mol. Biol. 175, 159- 174] and Fronticelli et al. [(1994) J. Biol. Chem. 269, 23965-23969] for deoxy-HbA. The electron density map of deoxy-DecHb indicates the presence of the 10 carbon bridge between the β82 lysines. Molecular modeling confirms that insertion of the linker into the T structure requires only slight displacement of the two β82 lysines. Instead, insertion of the linker into the R and R2 structures [Shaanan (1983) J. Mol. Biol. 171, 31-59; Silva et al. (1992) J. Biol. Chem. 267, 17248-17256] is hindered by serious sterical restrictions. The linker primarily affects the partially and fully liganded states of hemoglobin. The data suggest in DecHb concerted conformational changes at each step of oxygenation.

Original languageEnglish
Pages (from-to)3418-3425
Number of pages8
JournalBiochemistry
Volume35
Issue number11
DOIs
StatePublished - 19 Mar 1996

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Oxygenation
Lysine
Hemoglobins
Carbon
Oxygen
Molecular modeling
Carrier concentration
Isotherms
Electrons
Atoms
Peptides
Crystals

Cite this

Bucci, Enrico ; Razynska, Anna ; Kwansa, Herman ; Gryczynski, Zygmunt ; Collins, John H. ; Fronticelli, Clara ; Unger, Ron ; Braxenthaler, Michael ; Moult, John ; Ji, Xinhua ; Gilliland, Gary. / Positive and negative cooperativities at subsequent steps of oxygenation regulate the allosteric behavior of multistate sebacylhemoglobin. In: Biochemistry. 1996 ; Vol. 35, No. 11. pp. 3418-3425.
@article{9ca193cce986484c8f6a58b2ae6af53e,
title = "Positive and negative cooperativities at subsequent steps of oxygenation regulate the allosteric behavior of multistate sebacylhemoglobin",
abstract = "Cross-linked human hemoglobin (HbA) is obtained by reaction with bis(3,5- dibromosalicyl) sebacate. Peptide maps and crystallographic analyses confirm the presence of the 10 carbon atom long sebacyl residue cross-linking the two β82 lysines of the β-cleft (DecHb). The Adair's constants, obtained from the oxygen binding isotherms, show that at the first step of oxygenation normal hemoglobin and DecHb have a very similar oxygen affinity. In DecHb negative binding cooperativity is present at the second step of oxygenation, which has an affinity 27 times lower than at the first step. Positive cooperativity is present at the third binding step, whose affinity is 380 times that of the second step. The fourth binding step shows a weak negative cooperativity with an affinity one-half that of the third step. Crystals of deoxy-DecHb diffracted to 1.9 {\AA} resolution. The resulting atomic coordinates are very similar to those of Fermi et al. [(1984) J. Mol. Biol. 175, 159- 174] and Fronticelli et al. [(1994) J. Biol. Chem. 269, 23965-23969] for deoxy-HbA. The electron density map of deoxy-DecHb indicates the presence of the 10 carbon bridge between the β82 lysines. Molecular modeling confirms that insertion of the linker into the T structure requires only slight displacement of the two β82 lysines. Instead, insertion of the linker into the R and R2 structures [Shaanan (1983) J. Mol. Biol. 171, 31-59; Silva et al. (1992) J. Biol. Chem. 267, 17248-17256] is hindered by serious sterical restrictions. The linker primarily affects the partially and fully liganded states of hemoglobin. The data suggest in DecHb concerted conformational changes at each step of oxygenation.",
author = "Enrico Bucci and Anna Razynska and Herman Kwansa and Zygmunt Gryczynski and Collins, {John H.} and Clara Fronticelli and Ron Unger and Michael Braxenthaler and John Moult and Xinhua Ji and Gary Gilliland",
year = "1996",
month = "3",
day = "19",
doi = "10.1021/bi952446b",
language = "English",
volume = "35",
pages = "3418--3425",
journal = "Biochemistry",
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Bucci, E, Razynska, A, Kwansa, H, Gryczynski, Z, Collins, JH, Fronticelli, C, Unger, R, Braxenthaler, M, Moult, J, Ji, X & Gilliland, G 1996, 'Positive and negative cooperativities at subsequent steps of oxygenation regulate the allosteric behavior of multistate sebacylhemoglobin', Biochemistry, vol. 35, no. 11, pp. 3418-3425. https://doi.org/10.1021/bi952446b

Positive and negative cooperativities at subsequent steps of oxygenation regulate the allosteric behavior of multistate sebacylhemoglobin. / Bucci, Enrico; Razynska, Anna; Kwansa, Herman; Gryczynski, Zygmunt; Collins, John H.; Fronticelli, Clara; Unger, Ron; Braxenthaler, Michael; Moult, John; Ji, Xinhua; Gilliland, Gary.

In: Biochemistry, Vol. 35, No. 11, 19.03.1996, p. 3418-3425.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Positive and negative cooperativities at subsequent steps of oxygenation regulate the allosteric behavior of multistate sebacylhemoglobin

AU - Bucci, Enrico

AU - Razynska, Anna

AU - Kwansa, Herman

AU - Gryczynski, Zygmunt

AU - Collins, John H.

AU - Fronticelli, Clara

AU - Unger, Ron

AU - Braxenthaler, Michael

AU - Moult, John

AU - Ji, Xinhua

AU - Gilliland, Gary

PY - 1996/3/19

Y1 - 1996/3/19

N2 - Cross-linked human hemoglobin (HbA) is obtained by reaction with bis(3,5- dibromosalicyl) sebacate. Peptide maps and crystallographic analyses confirm the presence of the 10 carbon atom long sebacyl residue cross-linking the two β82 lysines of the β-cleft (DecHb). The Adair's constants, obtained from the oxygen binding isotherms, show that at the first step of oxygenation normal hemoglobin and DecHb have a very similar oxygen affinity. In DecHb negative binding cooperativity is present at the second step of oxygenation, which has an affinity 27 times lower than at the first step. Positive cooperativity is present at the third binding step, whose affinity is 380 times that of the second step. The fourth binding step shows a weak negative cooperativity with an affinity one-half that of the third step. Crystals of deoxy-DecHb diffracted to 1.9 Å resolution. The resulting atomic coordinates are very similar to those of Fermi et al. [(1984) J. Mol. Biol. 175, 159- 174] and Fronticelli et al. [(1994) J. Biol. Chem. 269, 23965-23969] for deoxy-HbA. The electron density map of deoxy-DecHb indicates the presence of the 10 carbon bridge between the β82 lysines. Molecular modeling confirms that insertion of the linker into the T structure requires only slight displacement of the two β82 lysines. Instead, insertion of the linker into the R and R2 structures [Shaanan (1983) J. Mol. Biol. 171, 31-59; Silva et al. (1992) J. Biol. Chem. 267, 17248-17256] is hindered by serious sterical restrictions. The linker primarily affects the partially and fully liganded states of hemoglobin. The data suggest in DecHb concerted conformational changes at each step of oxygenation.

AB - Cross-linked human hemoglobin (HbA) is obtained by reaction with bis(3,5- dibromosalicyl) sebacate. Peptide maps and crystallographic analyses confirm the presence of the 10 carbon atom long sebacyl residue cross-linking the two β82 lysines of the β-cleft (DecHb). The Adair's constants, obtained from the oxygen binding isotherms, show that at the first step of oxygenation normal hemoglobin and DecHb have a very similar oxygen affinity. In DecHb negative binding cooperativity is present at the second step of oxygenation, which has an affinity 27 times lower than at the first step. Positive cooperativity is present at the third binding step, whose affinity is 380 times that of the second step. The fourth binding step shows a weak negative cooperativity with an affinity one-half that of the third step. Crystals of deoxy-DecHb diffracted to 1.9 Å resolution. The resulting atomic coordinates are very similar to those of Fermi et al. [(1984) J. Mol. Biol. 175, 159- 174] and Fronticelli et al. [(1994) J. Biol. Chem. 269, 23965-23969] for deoxy-HbA. The electron density map of deoxy-DecHb indicates the presence of the 10 carbon bridge between the β82 lysines. Molecular modeling confirms that insertion of the linker into the T structure requires only slight displacement of the two β82 lysines. Instead, insertion of the linker into the R and R2 structures [Shaanan (1983) J. Mol. Biol. 171, 31-59; Silva et al. (1992) J. Biol. Chem. 267, 17248-17256] is hindered by serious sterical restrictions. The linker primarily affects the partially and fully liganded states of hemoglobin. The data suggest in DecHb concerted conformational changes at each step of oxygenation.

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U2 - 10.1021/bi952446b

DO - 10.1021/bi952446b

M3 - Article

C2 - 8639491

AN - SCOPUS:0040267932

VL - 35

SP - 3418

EP - 3425

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 11

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