Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2001 Mar 28;123(12):2929-30.
doi: 10.1021/ja0056668.

Concerted evolution of structure and function in a miniature protein

J W Chin, A Schepartz

Concerted evolution of structure and function in a miniature protein

J W Chin et al. J Am Chem Soc. .
No abstract available

PubMed Disclaimer

Figures

Figure 1
Figure 1
(a) Residues of PPBR4 targeted for variation mapped onto the crystal structure of aPP.4 Side chains varied in library A are in yellow, those varied in library B are in green. (b) Sequences of PPBR4 and the two libraries. Residues varied are indicated by an X. Each position was randomized at the DNA level using the NNS codon scheme. (c) Sequences of the N-terminal amino acids deduced from the DNA sequences of library B clones after three selection rounds. Peptides containing the boxed sequences followed by the remaining residues of PPBR4 were synthesized and their properties investigated in vitro.
Figure 2
Figure 2
(a & b) Plots illustrating the fraction of hsCRE bound (Θ) as a function of p011 (formula image), p012 (●), p016 (formula image), p007 (formula image), PPBR4 (formula image), and G27 (formula image) concentration at 4 °C. Each point represents the average of at least three trials. Error bars show the standard error. (c) Autoradiogram and plot illustrating the fraction of hsCRE bound (Θ) by p007 at 25 °C. (d) Plot illustrating the relative affinity of p007 for γ-[32P] hsCRE versus calf thymus DNA. The molar concentration of competing binding sites on calf thymus DNA (C) was estimated from the concentration of DNA in mg/mL and the molecular weight of a base pair assuming that every base on either strand of DNA represents the start of a competitor site. The inset shows the affinity of p007 for four mismatched duplexes that differ from hsCRE at 2 of 5 base pairs. Binding reactions were performed and data analyzed as described.,
Figure 3
Figure 3
Residues 1–31 of the minimized mean structure of p007 superimposed on the corresponding residues of aPP.
See this image and copyright information in PMC

References

    1. Zondlo NJ, Schepartz A. J. Am. Chem. Soc. 1999;121:6938. For earlier, successful efforts to design miniature, functional proteins, see references and .

    1. Cunningham BC, Wells JA. Curr. Op. Str. Biol. 1997;7:457. - PubMed
    2. Braisted AC, Wells JA. Proc. Natl. Acad. Sci., USA. 1996;93:5688. - PMC - PubMed
    1. Vita C, Roumestand C, Toma F, Menez A. Proc. Natl. Acad. USA. 1995;92:6404. - PMC - PubMed
    2. Lombardi A, Bryson JW, Ghirlanda G, DeGrado WF. J. Am. Chem. Soc. 1997;119:12378.
    3. Vita C, et al. Proc. Natl. Acad. USA. 1999;96:13091. - PMC - PubMed
    4. McColl DJ, Honchell CD, Frankel AD. Proc. Natl. Acad. USA. 1999;96:9521. - PMC - PubMed
    5. Domingues H, Cregut D, Sebald W, Oschkinat H, Serrano L. Nat. Str. Biol. 1999;6:652. - PubMed
    6. Segal DJ, Barbas CF., III Curr. Op. Chem. Biol. 2000;4:34. - PubMed
    1. Blundell TL, Pitts JE, Tickle IC, Wood SP, Wu C-W. Proc. Natl. Acad. Sci. USA. 1981;78:4175. - PMC - PubMed
    1. Zondlo N. Ph.D. thesis. Yale University; 1999.

LinkOut - more resources