Domain closure in mitochondrial aspartate aminotransferase

Catherine A. McPhalen, Michael G. Vincent, Daniel Picot, Johan N. Jansonius, Arthur Lesk, Cyrus Chothia

Research output: Contribution to journalArticle

157 Citations (Scopus)

Abstract

The subunits of the dimeric enzyme aspartate aminotransferase have two domains: one large and one small. The active site lies in a cavity that is close to both the subunit interface and the interface between the two domains. On binding the substrate the domains close together. This closure completely buries the substrate in the active site and moves two arginine side-chains so they form salt bridges with carboxylate groups of the substrate. The salt bridges hold the substrate close to the pyridoxal 5′-phosphate cofactor and in the right position and orientation for the catalysis of the transamination reaction. We describe here the structural changes that produce the domain movements and the closure of the active site. Structural changes occur at the interface between the domains and within the small domain itself. On closure, the core of the small domain rotates by 13 ° relative to the large domain. Two other regions of the small domain, which form part of the active site, move somewhat differently. A loop, residues 39 to 49, above the active site moves about 1 Å less than the core of the small domain. A helix within the small domain forms the "door" of the active site. It moves with the core of the small domain and, in addition, shifts by 1·2 Å, rotates by 10 °, and switches its first turn from the α to the 310 conformation. This results in the helix closing the active site. The domain movements are produced by a co-ordinated series of small changes. Within one subunit the polypeptide chain passes twice between the large and small domains. One link involves a peptide in an extended conformation. The second link is in the middle of a long helix that spans both domains. At the interface this helix is kinked and, on closure, the angle of the kink changes to accommodate the movement of the small domain. The interface between the domains is formed by 15 residues in the large domain packing against 12 residues in the small domain and the manner in which these residues pack is essentially the same in the open and closed structures. Domain movements involve changes in the main-chain and side-chain torsion angles in the residues on both sides of the interface. Most of these changes are small; only a few side-chains switch to new conformations. The result of these changes in conformation is to move interface residues in the small domain by between 0·2 and 3·3 Å relative to the large domain and so produce the relative rotation of the two domains. In both the open and closed forms the interface residues are close packed and almost all have normally allowed conformations.

Original languageEnglish (US)
Pages (from-to)197-213
Number of pages17
JournalJournal of Molecular Biology
Volume227
Issue number1
DOIs
StatePublished - Sep 5 1992

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Mitochondrial Aspartate Aminotransferase
Catalytic Domain
Salts
Peptides
Pyridoxal Phosphate
Aspartate Aminotransferases
Catalysis
Arginine

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Cite this

McPhalen, C. A., Vincent, M. G., Picot, D., Jansonius, J. N., Lesk, A., & Chothia, C. (1992). Domain closure in mitochondrial aspartate aminotransferase. Journal of Molecular Biology, 227(1), 197-213. https://doi.org/10.1016/0022-2836(92)90691-C
McPhalen, Catherine A. ; Vincent, Michael G. ; Picot, Daniel ; Jansonius, Johan N. ; Lesk, Arthur ; Chothia, Cyrus. / Domain closure in mitochondrial aspartate aminotransferase. In: Journal of Molecular Biology. 1992 ; Vol. 227, No. 1. pp. 197-213.
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McPhalen, CA, Vincent, MG, Picot, D, Jansonius, JN, Lesk, A & Chothia, C 1992, 'Domain closure in mitochondrial aspartate aminotransferase', Journal of Molecular Biology, vol. 227, no. 1, pp. 197-213. https://doi.org/10.1016/0022-2836(92)90691-C

Domain closure in mitochondrial aspartate aminotransferase. / McPhalen, Catherine A.; Vincent, Michael G.; Picot, Daniel; Jansonius, Johan N.; Lesk, Arthur; Chothia, Cyrus.

In: Journal of Molecular Biology, Vol. 227, No. 1, 05.09.1992, p. 197-213.

Research output: Contribution to journalArticle

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T1 - Domain closure in mitochondrial aspartate aminotransferase

AU - McPhalen, Catherine A.

AU - Vincent, Michael G.

AU - Picot, Daniel

AU - Jansonius, Johan N.

AU - Lesk, Arthur

AU - Chothia, Cyrus

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Y1 - 1992/9/5

N2 - The subunits of the dimeric enzyme aspartate aminotransferase have two domains: one large and one small. The active site lies in a cavity that is close to both the subunit interface and the interface between the two domains. On binding the substrate the domains close together. This closure completely buries the substrate in the active site and moves two arginine side-chains so they form salt bridges with carboxylate groups of the substrate. The salt bridges hold the substrate close to the pyridoxal 5′-phosphate cofactor and in the right position and orientation for the catalysis of the transamination reaction. We describe here the structural changes that produce the domain movements and the closure of the active site. Structural changes occur at the interface between the domains and within the small domain itself. On closure, the core of the small domain rotates by 13 ° relative to the large domain. Two other regions of the small domain, which form part of the active site, move somewhat differently. A loop, residues 39 to 49, above the active site moves about 1 Å less than the core of the small domain. A helix within the small domain forms the "door" of the active site. It moves with the core of the small domain and, in addition, shifts by 1·2 Å, rotates by 10 °, and switches its first turn from the α to the 310 conformation. This results in the helix closing the active site. The domain movements are produced by a co-ordinated series of small changes. Within one subunit the polypeptide chain passes twice between the large and small domains. One link involves a peptide in an extended conformation. The second link is in the middle of a long helix that spans both domains. At the interface this helix is kinked and, on closure, the angle of the kink changes to accommodate the movement of the small domain. The interface between the domains is formed by 15 residues in the large domain packing against 12 residues in the small domain and the manner in which these residues pack is essentially the same in the open and closed structures. Domain movements involve changes in the main-chain and side-chain torsion angles in the residues on both sides of the interface. Most of these changes are small; only a few side-chains switch to new conformations. The result of these changes in conformation is to move interface residues in the small domain by between 0·2 and 3·3 Å relative to the large domain and so produce the relative rotation of the two domains. In both the open and closed forms the interface residues are close packed and almost all have normally allowed conformations.

AB - The subunits of the dimeric enzyme aspartate aminotransferase have two domains: one large and one small. The active site lies in a cavity that is close to both the subunit interface and the interface between the two domains. On binding the substrate the domains close together. This closure completely buries the substrate in the active site and moves two arginine side-chains so they form salt bridges with carboxylate groups of the substrate. The salt bridges hold the substrate close to the pyridoxal 5′-phosphate cofactor and in the right position and orientation for the catalysis of the transamination reaction. We describe here the structural changes that produce the domain movements and the closure of the active site. Structural changes occur at the interface between the domains and within the small domain itself. On closure, the core of the small domain rotates by 13 ° relative to the large domain. Two other regions of the small domain, which form part of the active site, move somewhat differently. A loop, residues 39 to 49, above the active site moves about 1 Å less than the core of the small domain. A helix within the small domain forms the "door" of the active site. It moves with the core of the small domain and, in addition, shifts by 1·2 Å, rotates by 10 °, and switches its first turn from the α to the 310 conformation. This results in the helix closing the active site. The domain movements are produced by a co-ordinated series of small changes. Within one subunit the polypeptide chain passes twice between the large and small domains. One link involves a peptide in an extended conformation. The second link is in the middle of a long helix that spans both domains. At the interface this helix is kinked and, on closure, the angle of the kink changes to accommodate the movement of the small domain. The interface between the domains is formed by 15 residues in the large domain packing against 12 residues in the small domain and the manner in which these residues pack is essentially the same in the open and closed structures. Domain movements involve changes in the main-chain and side-chain torsion angles in the residues on both sides of the interface. Most of these changes are small; only a few side-chains switch to new conformations. The result of these changes in conformation is to move interface residues in the small domain by between 0·2 and 3·3 Å relative to the large domain and so produce the relative rotation of the two domains. In both the open and closed forms the interface residues are close packed and almost all have normally allowed conformations.

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