The structure of carboxypeptidase A

Lipscomb, William N.; Coppola, J. C.; Hartsuck, Jean A.; Ludwig, Martha; Muirhead, H.; Searl, James E.; Steitz, Thomas A.

doi:10.1016/s0022-2836(66)80014-1

Cited by 96 publications

(60 citation statements)

References 41 publications

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“…Comparisons with model building and biochemical data suggest that this is a poor substrate because it is bound in a way which interferes with the normal ftmction of the glutamic acid residue at position 270. lt is also suggested that the interaction between the carbonyl group of the substrate and the zinc ion is part of the normal catalytic mechanism, In fact, Lipscomb et al 32 have suggested a very plausible reaction scheme which is consistent with all available information, but for a discussion of this the reader is referred to the original paper. I would only like to stress that, while with both enzymes an essential role for the zinc ion in the catalytic mechanism is depicted, this function has no direct counterpart in the catalytic activity of simple zinc complexes.…”

Section: State and Function Of Metal Ions In Carboxypeptidase And Carmentioning

confidence: 86%

“…This view was enforced by a critical evaluation of the experimental carboxypeptidase papers, since, as I expressed in another review 30 , this shows that 'none of the experiments provides adefinite demonstration of the nature of the binding site'. Fortunately it has been possible to settle the ensuing conflict definitively, as the three-dimensional structure is now available both for carbonic anhydrase 31 and for carboxypeptidase 32 .…”

Section: State and Function Of Metal Ions In Carboxypeptidase And Carmentioning

confidence: 99%

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The 'unique' metal-binding properties of metalloenzymes

Malmström¹

1970

Pure and Applied Chemistry

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ABSTRACT(1) The theme of the survey is that metalloenzymes represent rather 'unique' metal complexes when compared to the simpler system generally studied by coordination chemists. This theme is introduced by a brief presentation of the historical development of the metalloenzyme field, particularly the contributions of 0. Warburg and D. Keilin.(2) Various types of metal ion-protein interactions and the distinction between non-specific and specific binding have been briefly reviewed.(3) The ·unique' properties in the specific interactions with metalloenzymes are illustrated by a discussion of three different enzymes: carboxypeptidase, carbonic anhydrase and laccase (p-diphenol oxidase). In conclusion it is suggested that in these enzymes the 'unique' coordination imparts properties to the metal ion facilitating its role in the catalytic reaction. METALLOENZYMES AND MODELSThe study of the role of metal ions in enzyme reactions provides a beautiful example of an area in which increased understanding has been intimately connected with the close interaction between traditionally distinct disciplines. Thus, the techniques and results of coordination chemistry, a field generally regarded as a branch of inorganic chemistry, have been utilized in investigations on one of the central problems of biochemistry, that of the relation between structure and function of enzymes. To my mind, a major impression from the progress made so far isthat metalloenzymes represent rather 'unique' metal complexes in comparison with the simpler systems generally studied by coordination chemists. The nature of these 'unique' metal-binding properties and their relation to the catalytic function of the metal ion in a number of metalloenzymes will form the main theme of this survey.The unusual coordination chemistry of metalloenzymes necessitates that arguments based on the properties of simpler 'model' complexes must be applied with caution and considerable judgement. This apparent antithesis between enzymes and models is weil illustrated by the different attitudes taken by two investigators, D. Keilin and 0. Warburg, who both played prominent roles in the early development of the field. Thus, it may be appropriate to introduce the subject by a brief account of some main points in the history of metalloenzymes.G. Bertrand was the first investigator who suggested a metal as the active group in enzymes. This occurred in the 1890s in connection with his work on

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Section: State and Function Of Metal Ions In Carboxypeptidase And Carmentioning

confidence: 86%

Section: State and Function Of Metal Ions In Carboxypeptidase And Carmentioning

confidence: 99%

The 'unique' metal-binding properties of metalloenzymes

Malmström¹

1970

Pure and Applied Chemistry

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“…However, identification of the endogenous cation using physico-chemical methods (X-ray, NMR, EPR) and analysis of the amino acid residues involved in the catalytic site should allow us to propose a hydrolysis mechanism of 5 '-AMP by 5'-nucleotidase, as in the case of carboxypeptidase A, a metalloprotein which requires zinc [21].…”

Section: Discussionmentioning

confidence: 99%

Evidence for a metalloprotein stucture of plasma membrane 5' ‐nucleotidase

Harb¹,

Méflah²,

Duflos³

et al. 1984

FEBS Letters

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To point out the metalloprotein structure of bovine liver plasma membrane 5 ' -nucleotidase, we studied the inhibition mechanism of the purified enzyme by EDTA: this apparently non-competitive inhibition seems to be dependent on EDTA concentration, pH, temperature and incubation time. When the restoration of activity was assayed by addition of divalent cations or by gel filtration, the inhibition became progressively irreversible with time. Incubation of the enzyme with [14C]EDTA allowed us to observe, after gel filtration as well as after sucrose gradient ultracentrifugation, that the chelating agent is bound to 5 ' -nucleotidase. '-NucleotidaseMetalloprotein

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“…So far, knowledge about the spatial conformation of the substrate binding regions and the catalytic mechanism of zinc-peptidases has stemmed from detailed kinetic experiments and X-ray structure analyses of carboxypeptidase A and thermolysin (Lipscomb et a]., 1968;Matthews et al, 1972;Holmquist and Vallee, 1976;Morgan and Fruton, 1978;Lipscomb, 1983;Vallee and Galdes, 1984;Auld et al, 1984;Auld, 1987;Matthews, 1988;Christiansen and Lipscomb, 1989). In these enzymes the active-site zinc is ligated by two histidine residues, a glutamic acid residue and a water molecule in a distorted tetrahedral coordination sphere.…”

Section: The Zinc-coordination Sitementioning

confidence: 99%

“…Although their overall structures are entirely different, the active sites of the endopeptidase thermolysin and of the exopeptidase carboxypeptidase A, have some properties in common (Matthews et al, 1972(Matthews et al, , 1974Lipscomb et al, 1968;Schmidt and Herriott, 1976). In both enzymes the central zinc ion is tetrahedrally complexed by two histidine residues, a glutamic acid residue and a glutamic-acid-bound water molecule.…”

mentioning

confidence: 99%

Implications of the three‐dimensional structure of astacin for the structure and function of the astacin family of zinc‐endopeptidases

Stöcker

Gomis-Rüth

Bode

et al. 1993

European Journal of Biochemistry

110

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Astacin, a zinc-endopeptidase from the crayfish Astacus astucus L., represents a structurally distinct group of metalloproteinases termed the 'astacin family'. This protein family includes oligomeric membrane-bound proteins with zinc proteinase domains found in rodent kidneys (meprins A and B) and human small intestine (N-benzoyl-~-tyrosyl-4-aminobenzoate hydrolase). Another branch of this family comprises morphogenetically active proteins, which induce bone formation (human bone morphogenetic protein 1), or which play specific roles during the embryonic development of amphibians, fishes, echinoderms, and insects.The X-ray crystal structure of astacin has recently been solved to a resolution of 0.18 nm [Bode et al. (1992) Nature 358, 164-1671. This structure is different from hitherto known metalloendopeptidase structures and has been used in the present study to analyze the structures of the other members of the astacin protein family.Computer-assisted modelling of the proteolytic domain of the a-subunit of meprin A based on the astacin structure is possible if five single and one double residue deletions and three single residue insertions are implied. The proteinase domains of the other astacins can be included in the model-based sequence alignment by introducing additionally three insertions and one deletion. All of these insertions and deletions are observed in loop segments connecting regular secondary structure elements and should leave the overall structure unaltered.The topology of residues forming the zinc-binding active site of astacin corresponds to almost identical arrangements in all other astacins, suggesting that these are likewise metalloproteinases. Based on this similarity, it is proposed that the active-site metal ion of the astacins is penta-coordinated by three histidine residues, a tyrosine residue and a water molecule in a trigonal bipyramidal geometry. Other remarkable common features are a hydrophobic cluster in the N-terminal domain and a conserved, solvent-filled cavity buried in the C-terminal domain. Most interestingly, the amino-termini of all astacins can be modelled to start in a corresponding internal water cavity as seen in the astacin template, where the terminal alanine residue forms a water-linked salt bridge to Glu103, directly adjacent to Hisl02, the third zinc ligand. Therefore, an activation mechanism for the astacins reminiscent of that of the trypsin-like proteinases had been suggested, which now seems to be probable also for the other astacins.Besides these common traits, there are some minor differences which may have important consequences on the function of the astacins. A striking example are variations in the presumed S: substrate-binding site, which binds the amino acid side chain on the C-terminal side of the scissile bond of the substrate. In this subsite the crayfish proteinase astacin prefers short, uncharged residues. By contrast, meprin A accepts bulky, charged side chains in this position. This difference presumably can be explained by both the replacemen...

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The structure of carboxypeptidase A

Cited by 96 publications

References 41 publications

The 'unique' metal-binding properties of metalloenzymes

The 'unique' metal-binding properties of metalloenzymes

Evidence for a metalloprotein stucture of plasma membrane 5' ‐nucleotidase

Implications of the three‐dimensional structure of astacin for the structure and function of the astacin family of zinc‐endopeptidases

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