Free energies of urea and of thermal unfolding show that two tandem repeats of spectrin are thermodynamically more stable than a single repeat - PubMed (original) (raw)
Comparative Study
. 2001 Apr 3;40(13):3974-84.
doi: 10.1021/bi0025159.
Affiliations
- PMID: 11300778
- DOI: 10.1021/bi0025159
Comparative Study
Free energies of urea and of thermal unfolding show that two tandem repeats of spectrin are thermodynamically more stable than a single repeat
R I MacDonald et al. Biochemistry. 2001.
Abstract
Free energies of both urea and thermal denaturation have been measured for three pairs of one- and two-repeat fragments, cloned in tandem from the cytoskeletal protein, alpha-spectrin, from chicken brain to ascertain whether one- and two-repeat fragments are equally stable. One- and two-repeat fragments of each pair were designed with the same N-terminus, whereas the C-terminus of the two-repeat fragment was 106 residues or the length of one repeat downstream from that of the one-repeat fragment. The averaged free energies of urea and thermal denaturation of the paired fragments, (R16)(00) and (R16R17)(00), (R16)(0+3) and (R16R17)(0+3), and (R16)(+8-4) and (R16R17)(+8-4) [subscripts represent the N- and C-terminal positions with "00" referring to the N- and C-termini defining a repeat according to X-ray crystal structures of two repeat fragments [Grum, V. L., Li, D., MacDonald, R. I., and Mondragón, A. (1999) Cell 98, 523-535] and "+" and "-" referring to positions upstream and downstream therefrom, respectively], increased from 3.7 +/- 0.4 kcal/mol for (R16)(00), 3.7 +/- 0.5 kcal/mol for (R16)(0+3), 4.4 +/- 0.4 kcal/mol for (R16)(+8-4), 6.2 +/- 0.6 kcal/mol for (R16R17)(+8-4), 8.3 +/- 0.4 kcal/mol for (R16R17)(00) to 9.9 +/- 1.0 kcal/mol for (R16R17)(0+3). Thus, the two-repeat fragment of each pair was significantly more thermodynamically stable than the single repeat by both urea and thermal denaturation. Differences in phasing among single repeats did not have the same effect as the same differences in phasing among two-repeat fragments. Addition of nine residues to the C-terminus of (R16R17)(00) yielded a free energy of unfolding of 7.9 +/- 0.8 kcal/mol, whereas addition of seven residues to the C-terminus of (R16)(+8-4) yielded a free energy of unfolding of 5.9 +/- 0.3 kcal/mol.
Similar articles
- Independent movement, dimerization and stability of tandem repeats of chicken brain alpha-spectrin.
Kusunoki H, Minasov G, Macdonald RI, Mondragón A. Kusunoki H, et al. J Mol Biol. 2004 Nov 19;344(2):495-511. doi: 10.1016/j.jmb.2004.09.019. J Mol Biol. 2004. PMID: 15522301 - Global analysis of the acid-induced and urea-induced unfolding of staphylococcal nuclease and two of its variants.
Ionescu RM, Eftink MR. Ionescu RM, et al. Biochemistry. 1997 Feb 4;36(5):1129-40. doi: 10.1021/bi9609681. Biochemistry. 1997. PMID: 9033404 - Urea and thermal equilibrium denaturation studies on the dimerization domain of Escherichia coli Trp repressor.
Gloss LM, Matthews CR. Gloss LM, et al. Biochemistry. 1997 May 13;36(19):5612-23. doi: 10.1021/bi970056e. Biochemistry. 1997. PMID: 9153401 - Thermal stability of chicken brain α-spectrin repeat 17: a spectroscopic study.
Brenner AK, Kieffer B, Travé G, Frøystein NA, Raae AJ. Brenner AK, et al. J Biomol NMR. 2012 Jun;53(2):71-83. doi: 10.1007/s10858-012-9620-y. Epub 2012 May 9. J Biomol NMR. 2012. PMID: 22569754
Cited by
- Functional Disassociation Between the Protein Domains of MSMEG_4305 of Mycolicibacterium smegmatis (Mycobacterium smegmatis) in vivo.
Czubat B, Minias A, Brzostek A, Żaczek A, Struś K, Zakrzewska-Czerwińska J, Dziadek J. Czubat B, et al. Front Microbiol. 2020 Aug 19;11:2008. doi: 10.3389/fmicb.2020.02008. eCollection 2020. Front Microbiol. 2020. PMID: 32973726 Free PMC article. - Cotranslational folding cooperativity of contiguous domains of α-spectrin.
Kemp G, Nilsson OB, Tian P, Best RB, von Heijne G. Kemp G, et al. Proc Natl Acad Sci U S A. 2020 Jun 23;117(25):14119-14126. doi: 10.1073/pnas.1909683117. Epub 2020 Jun 8. Proc Natl Acad Sci U S A. 2020. PMID: 32513720 Free PMC article. - Unraveling the Mechanical Unfolding Pathways of a Multidomain Protein: Phosphoglycerate Kinase.
Li Q, Scholl ZN, Marszalek PE. Li Q, et al. Biophys J. 2018 Jul 3;115(1):46-58. doi: 10.1016/j.bpj.2018.05.028. Biophys J. 2018. PMID: 29972811 Free PMC article. - Probing conformational stability and dynamics of erythroid and nonerythroid spectrin: effects of urea and guanidine hydrochloride.
Patra M, Mukhopadhyay C, Chakrabarti A. Patra M, et al. PLoS One. 2015 Jan 24;10(1):e0116991. doi: 10.1371/journal.pone.0116991. eCollection 2015. PLoS One. 2015. PMID: 25617632 Free PMC article. - Mechanism of assembly of the non-covalent spectrin tetramerization domain from intrinsically disordered partners.
Hill SA, Kwa LG, Shammas SL, Lee JC, Clarke J. Hill SA, et al. J Mol Biol. 2014 Jan 9;426(1):21-35. doi: 10.1016/j.jmb.2013.08.027. Epub 2013 Sep 17. J Mol Biol. 2014. PMID: 24055379 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources