Abstract
Optimal application of protein engineering technology will require understanding how the amino acid sequence of a polypeptide chain specifies its spatial conformation. It is now clear that polypeptide chain folding and subunit assembly proceeds through sequential pathways involving defined intermediates. The critical amino acid instructions directing these pathways appear to be dispersed through the sequence. Recent genetic approaches have begun to identify which residues in a chain are important in maintaining the pathway, and avoiding aggregated states. Investigations of the P22 tailspike endorhamnosidase suggest that one general class of mutants, tss mutants, may be particularly informative in identifying sequences determining the conformation of the intermediates. Certain inherited human diseases may represent single amino acid substitutions causing folding defects.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Kleid, D.G., Yansura, D., Small, B., Dowbenko, D., Moore, D.M., Grubman, M.J., McKercher, P.D., Morgan, D.O., Robertson, B.H. and Bachrach, H.L. 1981. Cloned viral protein vaccine for Foot-and-Mouth disease: Responses in cattle and swine. Science 214:1125–1129.
Simons, G., Remaut, E., Allet, B., Devos, R. and Fiers, W. 1984. High-level expression of human interferon gamma in Escherichia coli under control of the p1 promoter of bacteriophage lambda genes. Gene 28:55–64.
Schoner, R.G., Ellis, L.F. and Schoner, B.E. 1985. Isolation and purification of protein granules from Escherichia coli cells overproducing bovine growth hormone. Bio/Technology 3:151–154.
Masui, Y., Mizuno, T. and Inouye, M. 1984. Novel high-level expression cloning vehicles: 104-fold amplification of Escherichia coli minor protein. Bio/Technology 3:81–85.
Benson, S.A., Hall, M.N. and Silhavy, T.J. 1985. Genetic analysis of protein export in Escherichia coli. Ann. Rev. Biochem. 54:101–134.
Rossman, M. and Argos, P. 1981. Protein folding. Ann. Rev. Biochem 50:497–532.
Dickerson, R.E. and Geis, I. 1983. Hemoglobin: Structure, Function, Evolution, and Pathology. Benjamin Cummings, Menlo Park, CA.
Krebs, H., Schmid, F.X. and Jaenicke, R. 1983. Folding of homologous proteins. The refolding of different ribonucleases is independent of sequence variations, proline content and glycosylation. J. Mol. Biol. 169:619–635.
Ackers, G.K. and Smith, F.R. 1985. Effects of site-specific amino acid modifications on protein interactions and biological function. Ann. Rev. Biochem. 54:597–629.
Craik, C.S., Largman, C., Fletcher, T., Roczniak, S., Barr, P.J., Fletterick, R. and Rutter, W.J. 1985. Redesigning trypsin: Alteration of substrate specificity. Science 228:291–297.
Hecht, M.H. and Sauer, R.T. 1985. Phage lambda represser revertants. Amino acid substitutions that restore activity to mutant proteins. J. Mol. Biol. 186:53–63.
Epstein, R.H., Bolle, A., Steinberg, E.M., Kellenberger, E., Boy de la Tour, E., Chevalley, R., Edgar, R.S., Susman, M., Denhardt, G.H. and Lielausis, A. 1963. Physiological studies of conditional lethal mutants of bacteriophage T4D. Cold Spring Harbor Symp. Quant. Biol. 28:375–394.
Miller, J.H., Coulondre, C., Hofer, M., Schmeissner, U., Sommer, H. and Schmitz, A. 1979. Generation of altered proteins by the suppression of nonsense mutations. J. Mol. Biol. 131:191–222.
Kabsch, W. and Sander, C. 1984. On the use of sequence homologies to predict protein structure: Identical pentapeptides can have completely different conformations. Proc. Nat. Acad. Sci. USA. 81:1075–1078.
Brems, D.N. and Baldwin, R.L. 1985. Protection of amide protons in folding intermediates of ribonuclease A measured by pH-pulse exchange curves. Biochem. 24:1689–1693.
Kuwajima, K., Hiraoka, Y., Ikeguchi, M. and Sugai, S. 1985. Comparison of the transient folding intermediates in lysozyme and alpha-lactalbumin. Biochem. 24:874–881.
Baldwin, R.L. 1975. Intermediates in protein folding reactions and the mechanism of protein folding. Ann. Rev. Biochem. 44:453–475.
Kim, P.S. and Baldwin, R.L. 1982. Specific intermediates in the folding reactions of small proteins and the mechanism of protein folding pathways. Ann. Rev. Biochem. 51:459–489.
Creighton, T.E. 1978. Experimental studies of protein folding and unfolding. Prog. in Biophys. and Mol. Biol. 33:231–298.
Creighton, T.E. and Goldenberg, D.P. 1984. Kinetic role of a metastable native-like two-disulphide species in the folding transition of bovine pancreatic trypsin inhibitor. J. Mol. Biol. 179:497–526.
Lin, S.H., Konishi, Y., Nall, B.T. and Scheraga, H.A. 1985. Influence of an extrinsic cross-link on the folding pathway of ribonuclease A. Kinetics of folding-unfolding. Biochem. 24:2680–2686.
Mitchinson, C. and Pain, R.H. 1985. Effects of sulphate and urea on the stability and reversible unfolding of beta-lactamase from Staphylococcus aureus. Implications for the folding pathway of beta-lactamase. J. Mol. Biol. 184:331–342.
Blond, S. and Goldberg, M.E. 1985. Kinetics and importance of the dimerization step in the folding pathway of the beta-2 subunit of Escherichia coli tryptophan synthase. J. Mol. Biol. 182:597–606.
Jaenicke, R. and Rudolph, R. 1983. What NAD-dependent dehydrogenases teach us about the folding and association of oligomeric proteins, p.62–90. In: Biological Oxidations. H. Sund and V. Ullrich (eds.). Springer-Verlag, Heidelberg.
Blundell, T. and Wood, S. 1982. The conformation, flexibility, and dynamics of polypeptide hormones. Ann. Rev. Biochem. 51:123–154.
Kaiser, E.T. and Kezdy, F.J. 1984. Amphiphilic secondary structure: Design of peptide hormones. Science 223:249–255.
Briggs, M.S. and Gierasch, L.M. 1986. Molecular mechanisms of protein secretion: The role of the signal sequence. Adv. Prot. Chem. 38:in Press.
Bierzynski, A., Kim, P.S. and Baldwin, R.L. 1982. A salt bridge stabilizes the helix formed by isolated C-peptide of RNase A. Proc. Nat. Acad. Sci. USA. 79:2470–2474.
Kim, P.S. and Baldwin, R.L. 1984. A helix stop signal in the isolated S-peptide of ribonuclease A. Nature 307:329–334.
Grutter, M.G., Hawkes, R.B. and Matthews, B. 1979. Molecular basis of thermostability in the lysozyme from bacteriophage T4. Nature 277:667–669.
Hawkes, R., Grutter, M.G. and Schellman, J. 1984. Thermodynamic stability and point mutations of bacteriophage T4 lysozyme. J. Mol. Biol. 175:195–212.
Alber, T. and Wozniak, J.A. 1985. Genetic screen for mutations that increase the thermal stability of phage T4 lysozyme. Proc. Nat. Acad. Sci. USA. 82:747–750.
Hecht, M.H., Sturtevant, J.M. and Sauer, R.T. 1984. Effect of single amino acid replacements on the thermal stability of the amino-terminal domain of phage lambda represser. Proc. Nat. Acad. Sci. USA. 81:5685–5689.
Shire, S.J., Bock, L., Ogez, J., Builder, S., Kleid, D. and Moore, D.M. 1984. Purification and immunogenicity of fusion VP1 protein of foot and mouth disease virus. Biochem. 23:6474–6480.
Villafranca, J.E., Howell, E.E., Voet, D.H., Strobel, M.S., Ogden, R.C., Abelson, J.N. and Kraut, J. 1983. Directed mutagenesis of dihydrofolate reductase. Science 222:782–788.
Fersht, A.R., Shi, J.P., Knill-Jones, J., Lowe, D.M., Wilkinson, A.J., Blow, D.M., Brick, P., Carter, P., Waye, M.M.Y. and Winter, G. 1985. Hydrogen bonding and biological specificity analyzed by protein engineering. Nature 314:235–238.
Creighton, T.E. and Dyckes, D.F. 1981. Refolding of S-methionyl basic pancreatic trypsin inhibitor. J. Mol. Biol. 146:375–387.
Beasty, A.M. and Matthews, C.R. 1985. Characterization of an early intermediate in the folding of the alpha subunit of tryptophan synthase by hydrogen exchange measurement. Biochem. 24:3547–3553.
Matthews, C.R., Chrisanti, M.M., Manz, J.T. and Gepner, G.L. 1983. Effect of a single amino acid substitution on the folding of the alpha subunit of tryptophan synthase. Biochem. 22:1445–1452.
Shortle, D. and Lin, B. 1985. Genetic analysis of Staphylococcal nuclease: identification of three intragenic “global” suppressors of nuclease-minus mutations. Genetics 110:539–555.
Koshland, D. and Botstein, D. 1982. Evidence for postranslational translocation of beta-lactamase across the bacterial inner membrane. Cell 30:893–902.
Craig, S., Hollecker, M., Creighton, T.E. and Pain, R.H. 1985. Single amino acid substitutions block a late step in the folding of beta-lactamase from Staphylococcus aureus. J. Mol. Biol. 186:681–687.
Berget, P.B. and Poteete, A.R. 1980. Structure and functions of the phage P22 tail protein. J. Virol. 34:234–243.
Sauer, R.T., Krovatin, W., Poteete, A.R. and Berget, P.B. 1982. Phage P22 tail protein: Gene and amino acid sequence. Biochemistry 21:5811–5815.
Goldenberg, D. and King, J. 1981. Temperature-sensitive mutants blocked in the folding or subunit assembly of the bacteriophage P22 tail spike protein. II. Active mutant proteins matured at 30°C. J. Mol. Biol. 145:633–651.
Thomas, G.L., Jr., Li, Y., Fuller, M.T. and King, J. 1982. Structural studies of P22 phage, precursor particles and proteins by laser Raman spectroscopy. 1982. Biochem. 80:3866–3878.
Goldenberg, D. and King, J. 1982. Trimeric intermediate in the in vivo folding and subunit assembly of the tail spike endorhamnosidase of bacteriophage P22. Proc. Natl. Acad. Sci. USA. 79:3403–3407.
Goldenberg, D.P., Berget, P.B. and King, J. 1982. Maturation of the tail spike endorhamnosidase of Salmonella phage P22. J. Biol. Chem. 257:7864–7871.
Smith, D.H. and King, J. 1981. Temperature-sensitive mutants blocked in the folding or subunit assembly of the bacteriophage P22 tail spike protein. III. Inactive polypeptide chains synthesized at 39°C. J. Mol. Biol. 145:653–676.
Smith, D.H., Berget, P.B. and King, J. 1980. Temperature-sensitive mutants blocked in the folding or subunit assembly of the bacteriophage P22 tail-spike protein. I. Fine-structure mapping. Genetics 96:331–352.
Sadler, J.R. and Novick, A. 1965. The properties of represser and the kinetics of its action. J. Mol. Biol. 12:305–327.
Goldenberg, D.P., Smith, D.H. and King, J. 1983. Genetic analysis of the folding pathway for the tail spike protein of phage P22. Proc. Natl. Acad. Sci. USA 80:7060–7064.
Yu, M.-H. and King, J. 1984. Single amino acid substitutions influencing the folding pathway of the phage P22 tail spike endorhamnosidase. Proc. Natl. Acad. Sci. USA. 81:6584–6588.
Harrison, S.C. and Durbin, R. 1985. Is there a single pathway for folding of a polypeptide chain? Proc. Nat. Acad. Sci. USA. 82:4028–4030.
Rose, G.L., Gierasch, L.M. and Smith, J. 1985. Turns in Peptides and Proteins. Adv. Prot. Chem. 37:1–109.
Muller, K. and Garel, J-R. 1984. Folding of aspartokinase-homoserine dehydrogenase I is dominated by tertiary interactions. Biochem. 23:655–660.
Light, A. 1985. Protein solubility, protein modifications and protein folding. Biotechniques 3:298–306.
Kornfeld, R. and Kornfeld, S. 1985. Assembly of asparagine-linked oligosaccharides. Ann. Rev. Biochem. 54:631–664.
Pless, D.D. and Lennarz, W.J. 1977. Enzymatic conversion of proteins to glycoproteins. Proc. Nat. Acad. Sci. USA. 74:134–138.
Lau, J.T.Y., Welply, J.K., Shenbagamurthy, P., Naider, F. and Lennarz, W.J. 1983. Substrate recognition by oligosaccharide transferase. J. Biol. Chem. 258:15255–15260.
Gibson, R., Schlesinger, S. and Kornfeld, S. 1979. The nonglycosylated glycoprotein of vesicular stomatitis virus is temperature-sensitive and undergoes intracellular aggregation at elevated temperatures. J. Biol. Chem. 254:3600–3607.
Carrell, R.W., Bathurst, I.C. and Brennan, S.D. 1984. The molecular pathology of human alpha-1-antitrypsin. Biochem. Soc. Symp. 49:55–66.
Prockop, D.J., Kivirikko, K.I., Tuderman, L. and Guzman, N.A. 1979. The biosynthesis of collagen and its disorders. New Eng. J. Med. 301:13–23.
Steinman, B., Rao, V.H., Vogel, A., Bruckner, P., Gitzelmann, R. and Byers, P.H. 1984. Cysteine in the triple-helical domain of one allelic product of the alpha1(I) gene of type I collagen produces a lethal form of osteogenesis imperfecta. J. Biol. Chem. 259:1129–1138.
Bonadio, J., Holbrook, K.A., Gelinas, R.E., Jacob, J. and Byers, P.H. 1985. Altered triple helical structure of type I procollagen in lethal perinatal osteogenesis imperfecta. J. Biol. Chem. 260:1734–1742.
Freedman, R.B. 1984. Native disulphide bond formation in protein biosynthesis: Evidence for the role of protein disulphide isomerase. Trends Biochem. Sci. 9:438–441.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
King, J. Genetic Analysis of Protein Folding Pathways. Nat Biotechnol 4, 297–303 (1986). https://doi.org/10.1038/nbt0486-297
Issue Date:
DOI: https://doi.org/10.1038/nbt0486-297