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PDBsum entry 1lnc

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Hydrolase (metalloprotease) PDB id
1lnc
Jmol
Contents
Protein chain
316 a.a. *
Ligands
VAL-LYS
DMS
Metals
_MN ×3
_CA ×2
Waters ×170
* Residue conservation analysis

References listed in PDB file
Key reference
Title Structural analysis of zinc substitutions in the active site of thermolysin.
Authors D.R.Holland, A.C.Hausrath, D.Juers, B.W.Matthews.
Ref. Protein Sci, 1995, 4, 1955-1965. [DOI no: 10.1002/pro.5560041001]
PubMed id 8535232
Abstract
Native thermolysin binds a single catalytically essential zinc ion that is tetrahedrally coordinated by three protein ligands and a water molecule. During catalysis the zinc ligation is thought to change from fourfold to fivefold. Substitution of the active-site zinc with Cd2+, Mn2+, Fe2+, and Co2+ alters the catalytic activity (Holmquist B, Vallee BL, 1974, J Biol Chem 249:4601-4607). Excess zinc inhibits the enzyme. To investigate the structural basis of these changes in activity, we have determined the structures of a series of metal-substituted thermolysins at 1.7-1.9 A resolution. The structure of the Co(2+)-substituted enzyme is shown to be very similar to that of wild type except that two solvent molecules are liganded to the metal at positions that are thought to be occupied by the two oxygens of the hydrated scissile peptide in the transition state. Thus, the enhanced activity toward some substrates of the cobalt-relative to the zinc-substituted enzyme may be due to enhanced stabilization of the transition state. The ability of Zn2+ and Co2+ to accept tetrahedral coordination in the Michaelis complex, as well as fivefold coordination in the transition state, may also contribute to their effectiveness in catalysis. The Cd(2+)- and Mn(2+)-substituted thermolysins display conformational changes that disrupt the active site to varying degrees and could explain the associated reduction of activity. The conformational changes involve not only the essential catalytic residue, Glu 143, but also concerted side-chain rotations in the adjacent residues Met 120 and Leu 144. Some of these side-chain movements are similar to adjustments that have been observed previously in association with the "hinge-bending" motion that is presumed to occur during catalysis by the zinc endoproteases. In the presence of excess zinc, a second zinc ion is observed to bind at His 231 within 3.2 A of the zinc bound to native thermolysin, explaining the inhibitory effect.
Secondary reference #1
Title Structural comparison suggests that thermolysin and related neutral proteases undergo hinge-Bending motion during catalysis.
Authors D.R.Holland, D.E.Tronrud, H.W.Pley, K.M.Flaherty, W.Stark, J.N.Jansonius, D.B.Mckay, B.W.Matthews.
Ref. Biochemistry, 1992, 31, 11310-11316. [DOI no: 10.1021/bi00161a008]
PubMed id 1445869
Full text Abstract
Secondary reference #2
Title Structure of thermolysin refined at 1.6 a resolution.
Authors M.A.Holmes, B.W.Matthews.
Ref. J Mol Biol, 1982, 160, 623-639. [DOI no: 10.1016/0022-2836(82)90319-9]
PubMed id 7175940
Full text Abstract
Figure 3.
FIG. 3. Conformational diagram for the backbone of thermolysin. Residues that are outside the ``allowed'' regions for a hard-sphere model are numbered.
Figure 4.
FIG. 4. Stereo diagram illustrating the apparent thermal motion of t,he thermolysin molecule. Larger circles correspond to residues with greater apparen motion. The radius of each c~wlr l\as obtained 1)~ taking the verage R value for all atoms in that residue, subtracting a constant value of 4.0 AZ (in order to make differences in apparent motion more obvious) and drawing the circle at t,hr SO'?; probabilit? level (Johson, 196.5).
The above figures are reproduced from the cited reference with permission from Elsevier
Secondary reference #3
Title Structure of a mercaptan-Thermolysin complex illustrates mode of inhibition of zinc proteases by substrate-Analogue mercaptans.
Authors A.F.Monzingo, B.W.Matthews.
Ref. Biochemistry, 1982, 21, 3390-3394. [DOI no: 10.1021/bi00257a022]
PubMed id 7052122
Full text Abstract
Secondary reference #4
Title Binding of hydroxamic acid inhibitors to crystalline thermolysin suggests a pentacoordinate zinc intermediate in catalysis.
Authors M.A.Holmes, B.W.Matthews.
Ref. Biochemistry, 1981, 20, 6912-6920. [DOI no: 10.1021/bi00527a026]
PubMed id 7317361
Full text Abstract
Secondary reference #5
Title Binding of the biproduct analog l-Benzylsuccinic acid to thermolysin determined by X-Ray crystallography.
Authors M.C.Bolognesi, B.W.Matthews.
Ref. J Biol Chem, 1979, 254, 634-639.
PubMed id 762086
Abstract
Secondary reference #6
Title Comparison of the structures of carboxypeptidase a and thermolysin.
Authors W.R.Kester, B.W.Matthews.
Ref. J Biol Chem, 1977, 252, 7704-7710.
PubMed id 914833
Abstract
Secondary reference #7
Title A crystallographic study of the complex of phosphoramidon with thermolysin. A model for the presumed catalytic transition state and for the binding of extended substances.
Authors L.H.Weaver, W.R.Kester, B.W.Matthews.
Ref. J Mol Biol, 1977, 114, 119-132.
PubMed id 909082
Abstract
Secondary reference #8
Title Crystallographic study of the binding of dipeptide inhibitors to thermolysin: implications for the mechanism of catalysis.
Authors W.R.Kester, B.W.Matthews.
Ref. Biochemistry, 1977, 16, 2506-2516. [DOI no: 10.1021/bi00630a030]
PubMed id 861218
Full text Abstract
Secondary reference #9
Title Role of calcium in the thermal stability of thermolysin.
Authors F.W.Dahlquist, J.W.Long, W.L.Bigbee.
Ref. Biochemistry, 1976, 15, 1103-1111. [DOI no: 10.1021/bi00650a024]
PubMed id 814920
Full text Abstract
Secondary reference #10
Title Evidence of homologous relationship between thermolysin and neutral protease a of bacillus subtilis
Authors P.L.Levy, M.K.Pangburn, Y.Burstein, L.H.Ericsson, H.Neurath, K.A.Walsh.
Ref. atlas of protein sequence, 1976, 5, 98.
Secondary reference #11
Title The conformation of thermolysin.
Authors B.W.Matthews, L.H.Weaver, W.R.Kester.
Ref. J Biol Chem, 1974, 249, 8030-8044.
PubMed id 4214815
Abstract
Secondary reference #12
Title Binding of lanthanide ions to thermolysin.
Authors B.W.Matthews, L.H.Weaver.
Ref. Biochemistry, 1974, 13, 1719-1725. [DOI no: 10.1021/bi00705a025]
PubMed id 4831359
Full text Abstract
Secondary reference #13
Title The structure of thermolysin: an electron density map at 2-3 a resolution.
Authors P.M.Colman, J.N.Jansonius, B.W.Matthews.
Ref. J Mol Biol, 1972, 70, 701-724. [DOI no: 10.1016/0022-2836(72)90569-4]
PubMed id 5083153
Full text Abstract
Figure 5.
Fra. 5. Schematic diagram, seen from above, of the optical comparator used to build to thermolyein model after Richards, 1968).
Figure 9.
FIG. 9. (a) Key showing tho position of the major binding sites of Llra heavy BWIUS used to determirw thn phase trnglaa fur L~IC therrnolysin elootron density map. and for the folluwing diEcmnoe maps. The calcum positions LIY dotcnninod from tho throo -dinmtional map BK+ indicated in this Figure by crosses. DA&IA, imnrcury acetic acd. (b) DifYeronce electron density bdween atrontillm-thnrmolynin and native themolysin. Tho mnoutmn of this and the following maps are 2-4 L% (c) DifGrence electron den&y betweau barium-lborxnolyuin au3 uetivo thermolyti. (d) Diffeence obctro density between dysprosium-thermolyain and native thomolysin.
The above figures are reproduced from the cited reference with permission from Elsevier
Secondary reference #14
Title Amino-Acid sequence of thermolysin
Authors K.Titani, M.A.Hermodson, L.H.Ericsson, K.A.Walsh, H.Neurath.
Ref. nature new biol, 1972, 238, 35.
Secondary reference #15
Title Three dimensional structure of thermolysin
Authors B.W.Matthews, J.N.Jansonius, P.M.Colman, B.P.Schoenborn, D.Duporque.
Ref. nature new biol, 1972, 238, 37.
Secondary reference #16
Title Structure of thermolysin
Authors B.W.Matthews, P.M.Colman, J.N.Jansonius, K.Titani, K.A.Walsh, H.Neurath.
Ref. nature new biol, 1972, 238, 41.
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