NMR solution structure of the C-terminal fragment 255-316 of thermolysin: a dimer formed by subunits having the native structure.


:The solution structure of the C-terminal fragment 255-316 of thermolysin has been determined by two-dimensional proton NMR methods. For this disulfide-free fragment there was a previous proposal according to which it would fold into a stable helical structure forming a dimer at concentrations above 0.06 mM. A complete assignment of the proton NMR resonances of the backbone and amino acid side chains of the fragment was first performed using standard sequential assignment methods. On the basis of 729 distance constraints derived from unambiguously assigned nuclear Overhauser effect (NOE) proton connectivities, the three-dimensional structure of a monomeric unit was then determined by using distance geometry and restrained molecular dynamic methods. The globular structure of fragment 255-316 of thermolysin in solution, composed of three helices, is largely coincident with that of the corresponding region in the crystallographic structure of intact thermolysin [Holmes, M. A., & Matthews, B. W. (1982) J. Mol. Biol. 160, 623-639]. This fact allowed identification as intersubunit of up to 52 NOE cross correlations, which were used to dock the two subunits into a symmetric dimer structure. The obtained dimeric structure served as the starting structure in a final restrained molecular dynamic calculation subjected to a total of 1562 distance constraints. In the resulting dimeric structure, the interface between the two subunits, of a marked hydrophobic character, coincides topologically with the one between the 255-316 fragment and the rest of the protein in the intact enzyme. The present work decisively shows that the thermolysin fragment 255-316 can attain a stable and nativelike structure independently of the rest of the polypeptide chain. Considering that the thermolysin molecule is constituted of two structural domains of equal size (residues 1-157 and 158-316), the results of this study show that autonomously folding units can be substantially smaller than entire domains.






Rico M,Jiménez MA,González C,De Filippis V,Fontana A




Has Abstract


1994-12-13 00:00:00












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    pub_type: 杂志文章


    authors: Dancheck B,Nairn AC,Peti W

    更新日期:2008-11-25 00:00:00

  • Phosphoinositide 3-kinase in rat liver nuclei.

    abstract::Biochemical and immunochemical data from the present investigation reveal the existence of a p85/p110 phosphoinositide 3-kinase (PI 3-kinase) in rat liver nuclei. 32P-Labeling of membrane phosphoinositides by incubating intact nuclei with [gamma-32P]ATP results in the formation of [32P]phosphatidyl-inositol 3,4, 5-tri...


    pub_type: 杂志文章


    authors: Lu PJ,Hsu AL,Wang DS,Yan HY,Yin HL,Chen CS

    更新日期:1998-04-21 00:00:00

  • Observation of multiple myosin subfragment 1-ADP-fluoroberyllate complexes by 19F NMR spectroscopy.

    abstract::Fluoroaluminate and fluoroberyllate are potent inhibitors of the ATPase activity of myosin. Inhibition requires the presence of ADP, and much evidence has accumulated to suggest that the tetrahedral fluoroaluminate and fluoroberyllate ions act as phosphate analogues, binding with high affinity at the active site in th...


    pub_type: 杂志文章


    authors: Henry GD,Maruta S,Ikebe M,Sykes BD

    更新日期:1993-10-05 00:00:00

  • NMR studies of DNA support the role of pre-existing minor groove variations in nucleosome indirect readout.

    abstract::We investigated how the intrinsic sequence-dependent properties probed via the phosphate linkages (BI ↔ BII equilibrium) influence the preferred shape of free DNA, and how this affects the nucleosome formation. First, this exploits NMR solution studies of four B-DNA dodecamers that together cover 39 base pairs of the ...


    pub_type: 杂志文章


    authors: Xu X,Ben Imeddourene A,Zargarian L,Foloppe N,Mauffret O,Hartmann B

    更新日期:2014-09-09 00:00:00

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    abstract::Static FTIR light-induced difference spectra have been recorded for reaction centers from Rhodopseudomonas viridis in the following charge-separated states: P+QA(-)-PQA, P+QB(-)-PQB, I(-)-I, I-QA(-)-IQA, and I-QA(2-)-IQA. A comparison of the I(-)-I difference spectra with the I-QA(-)-IQA difference spectra reveals new...


    pub_type: 杂志文章


    authors: Buchanan S,Michel H,Gerwert K

    更新日期:1992-02-11 00:00:00

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    abstract::The mechanism of binding of the substrates Mg x ATP and ribose 5-phosphate as well as Mg2+ to the enzyme 5-phospho-D-ribosyl (alpha-1-diphosphate synthetase from Escherichia coli has been analyzed. By use of the competive inhibitors of ATP and ribose 5-phosphate binding, alpha,beta-methylene ATP and (+)-1-alpha,2-alph...


    pub_type: 杂志文章


    authors: Willemoës M,Hove-Jensen B

    更新日期:1997-04-22 00:00:00

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    pub_type: 杂志文章


    authors: Lieser SA,Davis TC,Helmann JD,Cohen SM

    更新日期:2003-11-04 00:00:00

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    pub_type: 杂志文章


    authors: Kosako H,Akamatsu Y,Tsurushita N,Lee KK,Gotoh Y,Nishida E

    更新日期:1996-10-08 00:00:00

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    authors: Burczyk M,Burkhalter MD,Blätte T,Matysik S,Caron MG,Barak LS,Philipp M

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    authors: Hou YX,Li CZ,Palaniyandi K,Magtibay PM,Homolya L,Sarkadi B,Chang XB

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    authors: Manceva S,Lin T,Pham H,Lewis JH,Goldman YE,Ostap EM

    更新日期:2007-10-23 00:00:00

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    authors: Annand RR,Kontoyianni M,Penzotti JE,Dudler T,Lybrand TP,Gelb MH

    更新日期:1996-04-09 00:00:00

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    authors: Zamparelli C,Ilari A,Verzili D,Giangiacomo L,Colotti G,Pascarella S,Chiancone E

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    pub_type: 杂志文章


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    pub_type: 杂志文章


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    更新日期:2006-03-21 00:00:00

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    pub_type: 杂志文章


    authors: Cai H,Strouse J,Dumlao D,Jung ME,Clarke S

    更新日期:2001-02-20 00:00:00

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    pub_type: 杂志文章


    authors: Williams DL Jr,Greene LE

    更新日期:1983-05-24 00:00:00