Correlation between sites of limited proteolysis and segmental mobility in thermolysin.

Limited proteolysis or autolysis of thermolysin under different experimental conditions leads to fission of a small number of peptide bonds located in exposed surface segments of the polypeptide chain characterized by highest mobility, as given by the temperature factors (B values) determined crystallographically [Holmes, M.A., & Matthews, B.W. (1982) J. Mol. Biol. 160, 623-639]. Considering also similar findings observed previously with other protein systems, it is proposed that this correlation between segmental mobility and sites of limited proteolysis in globular proteins is quite general. Thus, flexibility of the polypeptide chain of a globular protein at the site of proteolytic attack promotes optimal binding and proper interaction with the active site of the protease. These findings emphasize that apparent thermal motion seen in protein crystals is relevant to motion in solution and appear to be of general significance in protein-protein recognition processes.

[1]  C. Vita,et al.  Isolation of large peptides derived by cyanogen bromide cleavage of thermolysin using fast protein liquid chromatography (FPLC). , 2009, International journal of peptide and protein research.

[2]  C. Vita,et al.  Limited proteolysis of thermolysin by subtilisin: isolation and characterization of a partially active enzyme derivative. , 1985, Biochemistry.

[3]  C. Vita,et al.  Folding of thermolysin fragments. Identification of the minimum size of a carboxyl-terminal fragment that can fold into a stable native-like structure. , 1985, Journal of molecular biology.

[4]  Arthur J. Olson,et al.  The reactivity of anti-peptide antibodies is a function of the atomic mobility of sites in a protein , 1984, Nature.

[5]  D. Moras,et al.  Correlation between segmental mobility and the location of antigenic determinants in proteins , 1984, Nature.

[6]  J M Thornton,et al.  Amino and carboxy-terminal regions in globular proteins. , 1983, Journal of molecular biology.

[7]  J. Andrew McCammon,et al.  The dynamic picture of protein structure , 1983 .

[8]  B. Matthews,et al.  Structure of thermolysin refined at 1.6 A resolution. , 1982, Journal of molecular biology.

[9]  Hans Frauenfelder,et al.  Temperature-dependent X-ray diffraction as a probe of protein structural dynamics , 1979, Nature.

[10]  D. Phillips,et al.  Crystallographic studies of the dynamic properties of lysozyme , 1979, Nature.

[11]  I. Chaiken,et al.  Conformational and immunochemical analysis of the cyanogen bromide fragments of thermolysin. , 1979, Biochemistry.

[12]  M. Sternberg,et al.  Dynamic information from protein crystallography. An analysis of temperature factors from refinement of the hen egg-white lysozyme structure. , 1979, Journal of molecular biology.

[13]  T. Takano,et al.  Structure of myoglobin refined at 2-0 A resolution. I. Crystallographic refinement of metmyoglobin from sperm whale. , 1977, Journal of molecular biology.

[14]  B. Matthews,et al.  The conformation of thermolysin. , 1974, The Journal of biological chemistry.

[15]  M. Pangburn,et al.  Affinity chromatography of thermolysin and of neutral proteases from B. subtilis. , 1973, Biochemical and biophysical research communications.

[16]  D. Wetlaufer.  Nucleation, rapid folding, and globular intrachain regions in proteins. , 1973, Proceedings of the National Academy of Sciences of the United States of America.

[17]  B. Matthews,et al.  The structure of thermolysin: an electron density map at 2-3 A resolution. , 1972, Journal of molecular biology.

[18]  B P Schoenborn,et al.  Three-dimensional structure of thermolysin. , 1972, Nature: New biology.

[19]  A. Fujishima,et al.  Electrochemical Photolysis of Water at a Semiconductor Electrode , 1972, Nature.

[20]  K. Titani,et al.  Amino acid sequence of thermolysin. Isolation and characterization of the fragments obtained by cleavage with cyanogen bromide. , 1972, Biochemistry.

[21]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[22]  H. Tsuzuki,et al.  Thermolysin: kinetic study with oligopeptides. , 1970, European journal of biochemistry.

[23]  A. Berger,et al.  Mapping the active site of papain with the aid of peptide substrates and inhibitors. , 1970, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[24]  B. Vallee,et al.  Thermolysin: a zinc metalloenzyme. , 1969, Biochemical and biophysical research communications.

[25]  J. Feder.  A spectrophotometric assay for neutral protease. , 1968, Biochemical and biophysical research communications.

[26]  E. Shaw,et al.  Chromatography of trypsin and its derivatives. Characterization of a new active form of bovine trypsin. , 1968, The Journal of biological chemistry.

[27]  R. Porter.  The hydrolysis of rabbit y-globulin and antibodies with crystalline papain. , 1959, The Biochemical journal.

[28]  F. Richards,et al.  The preparation of subtilisn-modified ribonuclease and the separation of the peptide and protein components. , 1959, The Journal of biological chemistry.

[29]  J. Harris,et al.  Studies on pituitary polypeptide hormones. II. The action of pepsin and subtilisin on beta-melanocyte-stimulating hormone. , 1959, The Biochemical journal.

[30]  W. Bennett,et al.  Structural and functional aspects of domain motions in proteins. , 1984, CRC critical reviews in biochemistry.

[31]  B. Keil.  Enzymic Cleavage of Proteins , 1982 .

[32]  F. Gurd,et al.  Motions in proteins. , 1979, Advances in protein chemistry.

[33]  G. Voordouw,et al.  The structural and functional roles of metal ions in thermolysin. , 1978, CRC critical reviews in biochemistry.

[34]  M. Ottesen.  Induction of biological activity by limited proteolysis. , 1967, Annual review of biochemistry.

[35]  K. Linderstrøm-Lang,et al.  Structure and enzymatic break-down of proteins. , 1950 .