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Covalent Linkage: II. Intramolecular Linkages

  • Rosa Uy
  • Finn Wold

Abstract

In this section on immobilized enzymes a consideration of covalent intramolecular linkages may appear somewhat irrelevant in that very little, if any, immobilization can be achieved by intramolecular cross-linking reactions. However, since most of the techniques and reagents used for intermolecular cross-link formation are the same as those used to form intramolecular bonds, it is safe to conclude that intramolecular linkages are natural side reactions of many of the immobilization reactions. This, in turn, means that in assessing the properties of an immobilized enzyme, one needs to consider the effects of intramolecular cross-links as well as the effects of the immobilization, and such considerations should therefore justify the inclusion of this chapter. A brief review of nature’s own intramolecular protein cross-links should provide convincing evidence that the cross-link effects indeed can be quite substantial. The most obvious natural cross-link is the disulfide bond, which in most globular proteins imparts unique permanence to the proper three-dimensional folded structure. Especially in cases where the native molecule is composed of multiple polypeptide chains linked by interchain disulfide bonds is the cross-link an essential feature of the structural integrity of the molecule. In the case of some of the structural proteins, multiple disulfide bonds contribute structural rigidity and inertness to the molecular complexes. Similarly, in the case of the very stable and metabolically inert proteins of connective tissue, collagen and elastin, a whole group of unique cross-links derived from oxidatively deaminated lysines and hydroxylysines contribute to the structural properties and undoubtedly also to the metabolic intractability of these proteins. In the formation of the final product of the blood-clotting cascade, the fibrin network is cross-linked by intra- and intermolecular amide bonds between lysine -amino groups and glutamic acid γ-carboxyl groups through the action of the enzyme transglutaminase (factor XIII in the series of plasma clotting factors). Again the role of the cross-links appears to be to fix and stabilize specific molecular conformations and associations. Since the transglutaminase enzyme now has been found to be present in other tissues as well as in the plasma, it seems reasonable to conclude that this cross-linking reaction must be important to processes other than just coagulation.

Keywords

Covalent Linkage Subunit Structure FEBS Letter Glycerol Kinase Hexamethylene Diisocyanate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Alazard, R., Bechet, J. J., Dupaix, A., and Yon, J., 1973, Inactivation of a-chymotrypsin by a bifunctional reagent, 2-bromomethyl-3,1-benzoxazin-4-one, Biochim. Biophys. Acta 309: 379.Google Scholar
  2. Beaven, G. H., and Gratzer, W. B., 1973, Modification of the enzymatic activity of trypsin by intramolecular cross-links, Int. J. Peptide Protein Res. 5: 215.CrossRefGoogle Scholar
  3. Bechet, J. J., Dupaix, A., and Yon, J., 1973, Inactivation of a-chymotrypsin by a bifunctional reagent, 3,4-dihydro-3,4-dibromo-6-bromomethylcoumarin, Eur. J. Biochem. 35: 527.CrossRefGoogle Scholar
  4. Brandenburg, D., 1972, Preparation of N,,A N,B2s adiopoylinsulin, an intramolecularly cross-linked derivative of beef insulin, Z. Physiol. Chem. 353: 869.CrossRefGoogle Scholar
  5. Busse, W. D., and Carpenter, F. H., 1974, Carbonyl bis(L-methionine p-nitrophenyl ester): a new reagent for the reversible intramolecular cross-linking of insulin, J. Amer. Chem. Soc. 96:5947.CrossRefGoogle Scholar
  6. Busse, W. D., Hansen, S. R., and Carpenter, F. H., 1974, Carbonyl bis(L-methionyl) insulin: a proinsulin analog which is convertible to insulin, J. Amer. Chem. Soc. 96: 5959.CrossRefGoogle Scholar
  7. Davies, G. E., and Kaplan, G. J., 1972, Use of a diimidoester cross-linking reagent to examine the subunit structure of rabbit muscle pyruvate kinase, Can. J. Biochem. Physiol. 50: 416.CrossRefGoogle Scholar
  8. Davies, G. E., and Stark, G. R., 1970, Use of dimethyl suberimidate, a cross-linking reagent in studying the subunit structure of oligomeric proteins, Proc. Natl. Acad. Sci. U.S. 66: 651.CrossRefGoogle Scholar
  9. Dutton, A., and Singer, A. J., 1975, Cross-linking and labeling of membrane proteins by transglutaminase catalyzed reactions, Proc. Natl. Acad. Sci. U.S. 72: 2568.CrossRefGoogle Scholar
  10. Folk, J. E., and Chung, S. I., 1973, Molecular and catalytic properties of transglutaminase, in: Adv. Enzymol. 38: 109.Google Scholar
  11. Givol, D., 1969, Inactivation of glyceraldehyde 3-phosphate dehydrogenase by a bifunctional reagent, FEBS Letters 5: 153.CrossRefGoogle Scholar
  12. Grow, T. E., and Fried, M., 1975, Lipoprotein geometry: I. Spatial relationships of human HDL apoproteins studied with a bifunctional reagent, Biochem. Biophys. Res. Commun. 66: 352.CrossRefGoogle Scholar
  13. Guire, P. E., 1975, Photoreactive carrier derivatives for immobilization of enzymes and ligands, Federation Proc. 34: 690.Google Scholar
  14. Hartman, F. C., and Wold, F., 1967, Cross-linking of bovine pancreatic ribonuclease A with dimethyl adipimidate, Biochemistry 6: 2439.CrossRefGoogle Scholar
  15. Hixson, S. H., and Hixson, S. S., 1975, Azidophenyl bromide, a versatile photolabile bifunctional reagent, reaction with glyceraldehyde 3-phosphate dehydrogenase, Biochemistry 14: 4251.CrossRefGoogle Scholar
  16. Hucho, F., and Changeux, J. P., 1973, Molecular weight and quaternary structure of the cholinergic receptor protein extracted by detergent from electrophorus electricus electric tissue, FEBS Letters 38: 11.CrossRefGoogle Scholar
  17. Hucho, F., and Janda, M., 1974, Investigation of the quaternary structure of beef liver glutamate dehydrogenase with bifunctional reagents, Biochem. Biophys. Res. Commun. 57: 1080.CrossRefGoogle Scholar
  18. Hunter, M. J., and Ludwig, M. L., 1962, The reaction of imidoesters with proteins and related small molecules, J. Amer. Chem. Soc. 84: 3491.CrossRefGoogle Scholar
  19. Husain, S. S., and Lowe, G., 1968, Evidence for histidine in the active site of papain, Biochem. J. 108: 855.Google Scholar
  20. Husain, S. S., and Lowe, G., 1970, The amino acid sequence around active site cysteine and histidine residues of stem bromelain, Biochem. J. 117: 341.Google Scholar
  21. Husain, S. S., Ferguson, J. B., and Fruton, J. S., 1971, Bifunctional inhibitors of pepsin, Proc. Natl. Acad. Sci. U.S. 68: 2765.CrossRefGoogle Scholar
  22. Jarabak, J., and Street, M. A., 1971, Studies on the soluble 17/3-hydroxysteroid dehydrogenase from human placenta: evidence for a subunit structure, Biochemistry 10: 3831.CrossRefGoogle Scholar
  23. Josephs, R., Eisenberg, H., and Reisler, E., 1973, Some properties of cross-linked polymers of glutamic dehydrogenase, Biochemistry 12: 4060.CrossRefGoogle Scholar
  24. Kohlaw, G., and Boatman, G., 1971, Cross-linking of Salmonella isopropylmalate synthase with dimethyl suberimidate: evidence for antagonistic effects of leucine and acetyl CoA on the quaternary structure, Biochem. Biophys. Res. Commun. 43: 741.CrossRefGoogle Scholar
  25. Lad, P. M., and Hammes, G. G., 1974, Physical and chemical properties of rabbit muscle phosphofructokinase cross-linked with dimethyl suberimidate, Biochemistry 13: 4530.CrossRefGoogle Scholar
  26. Laursen, R. A. (ed.), 1975, Solid Phase Methods in Protein Sequence Analysis, Pierce Chemical Company, Rockford, Ill.Google Scholar
  27. Lindsay, D. G., 1971, Intramolecular cross-linked insulin, FEBS Letters 21: 105.CrossRefGoogle Scholar
  28. Lockhart, W. L., and Smith, D. B., 1975, Cross-linking of hemoglobin, haptoglobin and hemoglobinhaptoglobin complex with bifunctional reagents, Can. J. Biochem. Physiol. 53: 861.CrossRefGoogle Scholar
  29. Lubin, B. H., Pena, V., Metzer, W. C., Bymun, E., Bradley, T. B., and Packer, L., 1975, Dimethyl adipimidate: a new antisickling agent, Proc. Natl. Acad. Sci. U.S. 72: 43.CrossRefGoogle Scholar
  30. Lutter, L. C., Ortanderl, F., and Fasold, H., 1974, The use of a new series of cleavable protein-crosslinkers on the E. coli Ribosome, FEBS Letters 48: 288.CrossRefGoogle Scholar
  31. Matthyssens, G. E., and Verheulpen, W., 1973, Preparation characterization and antigenic specificity of a tyrosine-lysine cross-linked hen egg white lysozyme derivative, FEBS Letters 35: 239.CrossRefGoogle Scholar
  32. Means, G. R., and Feeney, R. E.. 1971, Chemical Modification of Proteins, Holden-Day, Inc., San Francisco.Google Scholar
  33. Olomucki, M., and Diopoh, J., 1972, Ethyl chloroacetimidate, its properties and its reaction with ribonuclease, Biochim. Biophys. Acta 263: 213.Google Scholar
  34. Ottesen, M., and Svensson, B., 1971, Modification of papain by treatment with glutaraldehyde under reducing and non-reducing conditions, Compt. Rend. Tray. Lab. Carlsberg 38: 171.Google Scholar
  35. Paul, R., and Anderson, G. W., 1960, N,N’-carbonyldiimidazole: a new peptide forming reagent, J. Amer. Chem. Soc. 82: 4596.CrossRefGoogle Scholar
  36. Ruoho, A., Bartlette, P. A., Dutton, A., and Singer S. J., 1975, A disulfide bridge bifunctional imidoester as a reversible cross-linking reagent, Biochem. Biophys. Res. Commun. 63: 417.CrossRefGoogle Scholar
  37. Shaltiel, S., and Tauber-Finkelstein, M., 1971, Introduction of an intramolecular cross-link at the active site of glyceraldehyde 3-phosphate dehydrogenase, Biochem. Biophys. Res. Commun. 44: 484.CrossRefGoogle Scholar
  38. Snyder, P. D., Jr., Wold, F., Bernlohr, R. W., Dullum, C., Desnick, R. J., Krivit, W., and Condie, R. M., 1974, Purified a-galactosidase A: stabilization to heat and protease degradation by complexing with antibody and by chemical modification, Biochim. Biophys. Acta 350: 432.Google Scholar
  39. Stach, R. W., and Shooter, E. M., 1974, The biological activity of cross-linked ß nerve growth factor protein, J. Biol. Chem. 249: 6668.Google Scholar
  40. Sun, T. T., Bollen, A., Kahan, L., and Traut, R. R., 1974, Topography of ribosomal proteins of E. coli 30S subunit as studied with the reversible cross-linking reagent methyl 4-mercaptobutyrimidate, Biochemistry 13: 2334.CrossRefGoogle Scholar
  41. Telford, J. N., Lad, P. M., and Hammes, G. G. 1975, Electron microscope study of native and cross-linked rabbit muscle phosphofructokinase, Proc. Natl. Acad. Sci. U.S. 72: 3054.CrossRefGoogle Scholar
  42. Thorner, J. W., and Paulus, H., 1971, Composition and subunit structure of glycerol kinase from E. coli, J. Biol. Chem. 246: 3885.Google Scholar
  43. Traut, R. R., Bollen, A., Sun, T. T., Hershey, J. W. B., Sunberg, J., and Pierce, L. R., 1973, Methyl 4mercaptobutyrimidate as a cleavable cross-linking reagent and its application to the E. coli ribosome, Biochemistry 12: 3266.CrossRefGoogle Scholar
  44. Van Driel, R., and Van Bruggen, E. F., 1975, Functional properties of them. modified hemocyanin: fixation of hemocyanin in the low and high oxygen affinity state by reaction with a bifunctional imido ester, Biochemistry 14: 730.CrossRefGoogle Scholar
  45. Wang, J. H. C., and Tu, J. I., 1969, Modification of glycogen phosphorylase b by glutaraldehyde: preparation and isolation of enzyme derivatives with enhanced stability, Biochemistry 8: 4403.CrossRefGoogle Scholar
  46. Wang, K., and Richards, F. M., 1974, An approach to nearest neighbor analysis of membrane proteins: application to the human erythrocyte membrane of a method employing cleavable cross-linkages, J. Biol. Chem. 249: 8005.Google Scholar
  47. Waterman, M. R., Yamaoka, K., Chuang, H. A., and Cottam, G. L., 1975, Anti-sickling nature of dimethyl adipimidate, Biochem. Biophys. Res. Commun. 63: 580.CrossRefGoogle Scholar
  48. Wold, F., 1972, Bifunctional Reagents, in: Methods in Enzymology (Vol. 23) ( C. H. W. Hirs and S. N. Timasheff, eds.), p. 623, Academic Press, New York.Google Scholar
  49. Wold, F., 1973, Chemical Modification of Enzymes, in: Enzyme Therapy in Genetic Diseases (Vol. IX, No. 2) ( D. Bergsma, ed.), p. 46, Williams and Wilkins, Baltimore.Google Scholar
  50. Young, R. A., and Blumenthal, T., 1975, Phage Q-B ribonucleic acid replicase subunit relationships determined by intramolecular cross-linking, J. Biol. Chem. 250: 1829.Google Scholar

Copyright information

© Plenum Press, New York 1977

Authors and Affiliations

  • Rosa Uy
    • 1
  • Finn Wold
    • 1
  1. 1.Department of BiochemistryUniversity of MinnesotaSt. PaulUSA

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