The design, synthesis, and crystallization of an alpha‐helical peptide

Twelve‐ and sixteen‐residue peptides have been designed to form tetrameric alphahelical bundles. Both peptides are capable of folding into amphiphilic alpha‐helices, with leucyl residues along one face and glutamyl and lysyl residues along the opposite face. Four such amphiphilic alpha‐helices are capable of forming a noncovalently bonded tetramer. Neighboring helices run in antiparallel directions in the design, so that the complex has 222 symmetry. In the designed tetramer, the leucyl side chains interdigitate in the center in a hydrophobic interaction, and charged side chains are exposed to the solvent. The designed 12‐mer(ALPHA‐1) has been synthesized, and it forms helical aggregates in aqueous solution as judged by circular dichroic spectroscopy. It has also been crystallized and characterized by x‐ray diffraction. The crystal symmetry is compatible with (but does not prove) the design. The design can be extended to a four‐alpha‐helical bundle formed from a single polypeptide by adding three peptide linkers.

[1]  C. Pace,et al.  Urea and Guanidine Hydrochloride Denaturation of Ribonuclease , Lysozyme , & Zhymotrypsin , and @ Lactoglobulin * , 2003 .

[2]  A. D. McLachlan,et al.  Solvation energy in protein folding and binding , 1986, Nature.

[3]  William F. DeGrado,et al.  Induction of peptide conformation at apolar water interfaces. 1. A study with model peptides of defined hydrophobic periodicity , 1985 .

[4]  M. Sundaralingam,et al.  Stabilization of the long central helix of troponin C by intrahelical salt bridges between charged amino acid side chains. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[5]  G. Rose,et al.  Hydrophobicity of amino acid residues in globular proteins. , 1985, Science.

[6]  M. Mutter.  The Construction of New Proteins and Enzymes‐a Prospect for the Future? , 1985 .

[7]  H. Guy.  Amino acid side-chain partition energies and distribution of residues in soluble proteins. , 1985, Biophysical journal.

[8]  R. Hodges,et al.  Synthesis of a model protein of defined secondary and quaternary structure. Effect of chain length on the stabilization and formation of two-stranded alpha-helical coiled-coils. , 1984, The Journal of biological chemistry.

[9]  Harold A. Scheraga,et al.  Helix-coil stability constants for the naturally occurring amino acids in water. 22. Histidine parameters from random poly[(hydroxybutyl)glutamine-co-L-histidine] , 1984 .

[10]  W. Kullmann.  Design, synthesis, and binding characteristics of an opiate receptor mimetic peptide. , 1984, Journal of medicinal chemistry.

[11]  E. Kaiser,et al.  Amphiphilic secondary structure: design of peptide hormones. , 1984, Science.

[12]  C. Chothia.  Principles that determine the structure of proteins. , 1984, Annual review of biochemistry.

[13]  D. Eisenberg,et al.  The hydrophobic moment detects periodicity in protein hydrophobicity. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Bernd Gutte,et al.  An artificial crystalline DDT‐binding polypeptide , 1983 .

[15]  C. Pabo.  Molecular technology: Designing proteins and peptides , 1983, Nature.

[16]  W. DeGrado,et al.  Solid-phase synthesis of protected peptides on a polymer-bound oxime: preparation of segments comprising the sequence of a cytotoxic 26-peptide analog , 1982 .

[17]  F R Salemme,et al.  alpha-Helix dipole model and electrostatic stabilization of 4-alpha-helical proteins. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[18]  D. Eisenberg,et al.  Hydrophobic moments and protein structure , 1982 .

[19]  Wim G. J. Hol,et al.  Dipoles of the α-helix and β-sheet: their role in protein folding , 1981, Nature.

[20]  A. Brack,et al.  Multiconformational synthetic polypeptides , 1981 .

[21]  F. R. Salemme,et al.  Structural and functional diversity in 4-α-helical proteins , 1980, Nature.

[22]  W. V. Shaw,et al.  The amino acid sequence of the delta haemolysin of Staphylococcus aureus , 1980, FEBS letters.

[23]  J. Janin,et al.  Surface and inside volumes in globular proteins , 1979, Nature.

[24]  H. Scheraga.  Use of random copolymers to determine the helix-coil stability constants of the naturally occurring amino acids , 1978 .

[25]  C. Chothia,et al.  Structure of proteins: packing of alpha-helices and pleated sheets. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[26]  M. Basu,et al.  Surface properties of membrane systems. Transport of staphylococcal delta-toxin from aqueous to membrane phase. , 1977, Biochimica et biophysica acta.

[27]  Ian J. Tickle,et al.  X-ray analysis of glucagon and its relationship to receptor binding , 1975, Nature.

[28]  H. Scheraga,et al.  The effect of neighboring charges on the helix forming ability of charged amino acids in proteins. , 1975, Macromolecules.

[29]  D. Blow,et al.  Letter: Crystallization and preliminary x-ray diffraction studies on tyrosyl-transfer RNA synthetase from Bacillus stearothermophilus. , 1973, Journal of molecular biology.

[30]  E. Kaiser,et al.  Color test for detection of free terminal amino groups in the solid-phase synthesis of peptides. , 1970, Analytical biochemistry.

[31]  G. Fasman,et al.  Computed circular dichroism spectra for the evaluation of protein conformation. , 1969, Biochemistry.