Three-dimensional Fourier Synthesis of Tosyl-elastase at 3.5 Å Resolution

Watson, Herman C.; Shotton, David M.; Cox, Joyce M.; Muirhead, H.

doi:10.1038/225806a0

Cited by 148 publications

(44 citation statements)

References 25 publications

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“…A modified Watson scan procedure [9] described in detail elsewhere [lo] was used in order to optimize accuracy and time in the intensity measurement of each individual reflexion. The intensities were corrected for Lorentz and polarization effects in the usual way.…”

Section: Methodsmentioning

confidence: 99%

The Crystal Structure of Complexes between Horse Liver Alcohol Dehydrogenase and the Coenzyme Analogues 3‐Iodopyridine‐adenine Dinucleotide and Pyridine‐adenine Dinucleotide

Samama

Zeppezauer

Biellmann

et al. 1977

European Journal of Biochemistry

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We have studied the binding of the enzymatically active NAD + analogue, 3-iodopyridine-adenine dinucleotide, and the inactive analogue, pyridine-adenine dinucleotide to the enzyme horse liver alcohol dehydrogenase using X-ray crystallographic methods. These studies were made under such conditions that crystals of the complexes were isomorphous to apocnzyme crystals. Both analogues bind in the same conformation. The binding of the adenosine moiety is very similar to that of ADP-ribose or NADH bound to the enzyme. The conformation and mode of binding of the remaining portions of the analogue molecules is, however, quite different. The pyridine ring is not situated in the active-site pocket as the nicotinamide group in thc isomorphous enzyme -NADH . imidazole complex but lies at the surface of the crevice between the two domains of the subunit, approximately 1.5 nm away from the catalytically active zinc atom. Lys-228 which has been shown to be important for NADH dissociation is in this region of the molecule.A large number of analogues to the coenzyme NAD' have been investigated [l] in order to delineate the role of the various moieties of the dinucleotide in the interaction of this coenzyme with the NAD+-dependent dehydrogenases. We have been particularly interested in the enzymatic effects of substitutions on the nicotinamide ring. As part of these studies, and also for thc use as heavy-atom derivatives in protein Crystallography, we synthesized 3-iOdO and subsequently 3-bromo and 3-chloro derivatives of pyridineadenine dinucleotide which were active in enzymatic hydrogen transfer reactions. Synthesis of these analogues and their kinetic properties for some NAD+-dependent dehydrogenase-catalysed reactions is presented elsewhere [2].The present study describes the binding of the 3-iodo analogue to one of these enzymes, horse liver alcohol dehydrogenase, as studied by X-ray crystallographic methods. Furthermore, the results of that Abbreviations.Pd AD + . pyridine-adenine dinucleot ide ; io3PdAD+, 3-iodopyridine-adenine dinucleotide; Tes, 2-([tris-(hydroxymethyl)methyi]amino)ethane sulfonic acid.

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Section: Methodsmentioning

confidence: 99%

The Crystal Structure of Complexes between Horse Liver Alcohol Dehydrogenase and the Coenzyme Analogues 3‐Iodopyridine‐adenine Dinucleotide and Pyridine‐adenine Dinucleotide

Samama

Zeppezauer

Biellmann

et al. 1977

European Journal of Biochemistry

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“…While a major contribution to the binding energy in chymotrypsin and trypsin probably arises from the perfect fit of the sidechain of P~ in the ,,pocket,, (S~), this will hardly be the case for subtilisin and elastase. In subtilisin the S~ is much more open than in chymotrypsin and trypsin, and in elastase the entrance to the ~,pocket,, is blocked by a Val (210). The best fit to S~ is obtained by Ala which therefore to a large extent determines the specificity of elastase.…”

Section: Elastasementioning

confidence: 88%

Chemical modifications of the subtilisins with special reference to the binding of large substrates. A review

Svendsen

1976

Carlsberg Res. Commun.

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Subtilisin type Carlsberg and subtilisin BPN' are two well characterized extracellular proteases from Bacillus subtilis and Bacillus amyloquefaciens, respectively. The present review summarizes the various chemical modification studies which have been performed with these enzymes. Of special interest has been those modifications which lead to changes in the enzymatic properties with regard to the hydrolysis of polypeptide substrates but not small synthetic ester substrates. In this way it is possible to obtain information about how large an area of the surface of the enzyme is involved in the binding of natural substrates (e.g. clupein, gelatine, casein). Nitration, iodination, glutarylation or succinylation of the subtilisins leads to increases in the hydrolyses of clupein and gelatine, while modification of carboxyl groups leads to a decrease. In all these cases the hydrolysis of small ester substrates is almost unaffected. A section of the review deals with the comparison between the results obtained by chemical modification studies and the two other methods available for the study of "secondary binding sites": X-ray diffraction analyses and kinetic studies with synthetic polypeptides. The productive binding mode for polypeptides proposed by KRAUT and coworkers is in agreement with the results obtained by nitration of the subtilisins. However, results from our laboratory and the literature point towards more than one mode of productive binding. These results are discussed. Finally a comparison is made between the secondary binding sites in subtilisin and other proteolytic enzymes, especially the serine proteases. The importance of secondary interactions between enzyme and substrate is discussed.

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“…The space groups were determined from photographs, and the unit-cell dimensions by a leastsquares refinement from the setting angles of several reflexions measured at both positive and negative 0 on a Hilger & Watts Y290 PDP8-controlled four-circle diffractometer. Intensity data were measured by an to/20 scan with ordinate analysis (Watson, Shotton, Cox & Muirhead, 1970) using Mo Ka radiation from a graphite monochromator. Reflexions with [ < 3o(/") or displaced by more than 0.15 ° from their predicted positions were eliminated from the data sets.…”

Section: Methodsmentioning

confidence: 99%

Structures of three cis-dioxoazacycloalkanes: 3,3,4,4-tetramethyldiazetine 1,2-dioxide, 2,3-diazabicyclo[2.2.1]hept-2-ene 2,3-dioxide and 6,7-diazatetracyclo[3.2.1.1^3,8.0^2,4]non-6-ene 6,7-dioxide

Prout¹,

Stothard²,

Watkin³

1978

Acta Crystallogr Sect B

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Three-dimensional Fourier Synthesis of Tosyl-elastase at 3.5 Å Resolution

Cited by 148 publications

References 25 publications

The Crystal Structure of Complexes between Horse Liver Alcohol Dehydrogenase and the Coenzyme Analogues 3‐Iodopyridine‐adenine Dinucleotide and Pyridine‐adenine Dinucleotide

The Crystal Structure of Complexes between Horse Liver Alcohol Dehydrogenase and the Coenzyme Analogues 3‐Iodopyridine‐adenine Dinucleotide and Pyridine‐adenine Dinucleotide

Chemical modifications of the subtilisins with special reference to the binding of large substrates. A review

Structures of three cis-dioxoazacycloalkanes: 3,3,4,4-tetramethyldiazetine 1,2-dioxide, 2,3-diazabicyclo[2.2.1]hept-2-ene 2,3-dioxide and 6,7-diazatetracyclo[3.2.1.1^3,8.0^2,4]non-6-ene 6,7-dioxide

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Three-dimensional Fourier Synthesis of Tosyl-elastase at 3.5 Å Resolution

Cited by 148 publications

References 25 publications

The Crystal Structure of Complexes between Horse Liver Alcohol Dehydrogenase and the Coenzyme Analogues 3‐Iodopyridine‐adenine Dinucleotide and Pyridine‐adenine Dinucleotide

The Crystal Structure of Complexes between Horse Liver Alcohol Dehydrogenase and the Coenzyme Analogues 3‐Iodopyridine‐adenine Dinucleotide and Pyridine‐adenine Dinucleotide

Chemical modifications of the subtilisins with special reference to the binding of large substrates. A review

Structures of three cis-dioxoazacycloalkanes: 3,3,4,4-tetramethyldiazetine 1,2-dioxide, 2,3-diazabicyclo[2.2.1]hept-2-ene 2,3-dioxide and 6,7-diazatetracyclo[3.2.1.13,8.02,4]non-6-ene 6,7-dioxide

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Structures of three cis-dioxoazacycloalkanes: 3,3,4,4-tetramethyldiazetine 1,2-dioxide, 2,3-diazabicyclo[2.2.1]hept-2-ene 2,3-dioxide and 6,7-diazatetracyclo[3.2.1.1^3,8.0^2,4]non-6-ene 6,7-dioxide