Four-helix bundle topology re-engineered: monomeric Rop protein variants with different loop arrangements.

We converted the small homodimeric four-helix bundle repressor of primer protein (Rop) into a monomeric four-helix bundle by introduction of connecting loops. Both left- and right-handed four-helix bundles were produced. The left-handed bundles were more stable and were used to introduce biologically interesting peptides in one of the loops.

[1]  L. Serrano,et al.  Reading protein sequences backwards. , 1998, Folding & design.

[2]  M. Helmer-Citterich,et al.  Control of ColE1 plasmid replication by antisense RNA. , 1991, Trends in genetics : TIG.

[3]  G J Williams,et al.  The Protein Data Bank: a computer-based archival file for macromolecular structures. , 1977, Journal of molecular biology.

[4]  W. DeGrado,et al.  Solution structure and dynamics of a de novo designed three-helix bundle protein. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[5]  W. DeGrado,et al.  Protein Design: A Hierarchic Approach , 1995, Science.

[6]  H. Schägger,et al.  Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. , 1987, Analytical biochemistry.

[7]  M Kokkinidis,et al.  Structure of the ColE1 rop protein at 1.7 A resolution. , 1987, Journal of molecular biology.

[8]  A Kolinski,et al.  Does a backwardly read protein sequence have a unique native state? , 1996, Protein engineering.

[9]  H. Fritz,et al.  Efficient oligonucleotide-directed construction of mutations in expression vectors by the gapped duplex DNA method using alternating selectable markers. , 1989, Nucleic acids research.

[10]  G Vriend,et al.  WHAT IF: a molecular modeling and drug design program. , 1990, Journal of molecular graphics.

[11]  S. Provencher CONTIN: A general purpose constrained regularization program for inverting noisy linear algebraic and integral equations , 1984 .

[12]  L Regan,et al.  Redesigning the topology of a four-helix-bundle protein: monomeric Rop. , 1995, Biochemistry.

[13]  D. J. Thomas The entropic tension of protein loops. , 1990, Journal of molecular biology.

[14]  Proton nuclear magnetic resonance assignments and secondary structure determination of the ColE1 rop (rom) protein. , 1990, Biochemistry.

[15]  C. Sander Design of protein structures: helix bundles and beyond. , 1994, Trends in biotechnology.

[16]  K. Kirschner,et al.  A fully active variant of dihydrofolate reductase with a circularly permuted sequence. , 1992, Biochemistry.

[17]  L Regan,et al.  An inverse correlation between loop length and stability in a four-helix-bundle protein. , 1997, Folding & design.

[18]  T. Alber,et al.  Circular permutation of T4 lysozyme. , 1993, Biochemistry.

[19]  L. Castagnoli,et al.  Control of initiation of pMB1 replication: Purified rop protein and RNA I affect primer formation in vitro , 1984, Cell.

[20]  G. Cesareni,et al.  Crystallization of the ColE1 Rop protein. , 1983, Journal of molecular biology.

[21]  A. Finkelstein,et al.  S6 permutein shows that the unusual target topology is not responsible for the absence of rigid tertiary structure in de novo protein albebetin , 1997, FEBS letters.

[22]  H. Blöcker,et al.  Simultaneous synthesis and biological applications of DNA fragments: an efficient and complete methodology. , 1987, Methods in enzymology.

[23]  L. Regan,et al.  Characterization of a helical protein designed from first principles. , 1988, Science.

[24]  T. Hynes,et al.  Transfer of a β-turn structure to a new protein context , 1989, Nature.

[25]  D. Hilvert,et al.  Redesigning enzyme topology by directed evolution. , 1998, Science.

[26]  G. Cesareni,et al.  Introduction of a Proline Residue into Position 31 of the Loop of the Dimeric 4-α-Helical Protein ROP Causes a Drastic Destabilization , 1997, Biological chemistry.

[27]  R. Huber,et al.  Crystallographic refinement of Japanese quail ovomucoid, a Kazal-type inhibitor, and model building studies of complexes with serine proteases. , 1982, Journal of molecular biology.

[28]  W G Hol,et al.  Structure of bovine liver rhodanese. I. Structure determination at 2.5 A resolution and a comparison of the conformation and sequence of its two domains. , 1978, Journal of Molecular Biology.

[29]  Udo Heinemann,et al.  Crystal structures and properties of de novo circularly permuted 1,3‐1,4‐β‐glucanases , 1998, Proteins.