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2let

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[[Image:2let.gif|left|200px]]
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{{STRUCTURE_2let| PDB=2let | SCENE= }}
{{STRUCTURE_2let| PDB=2let | SCENE= }}
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'''AN 1H NMR DETERMINATION OF THE THREE DIMENSIONAL STRUCTURES OF MIRROR IMAGE FORMS OF A LEU-5 VARIANT OF THE TRYPSIN INHIBITOR ECBALLIUM ELATERIUM (EETI-II)'''
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===AN 1H NMR DETERMINATION OF THE THREE DIMENSIONAL STRUCTURES OF MIRROR IMAGE FORMS OF A LEU-5 VARIANT OF THE TRYPSIN INHIBITOR ECBALLIUM ELATERIUM (EETI-II)===
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==Overview==
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The 3-dimensional structures of mirror-image forms of a Leu-5 variant of the trypsin inhibitor Ecballium elaterium (EETI-II) have been determined by 1H NMR spectroscopy and simulated annealing calculations incorporating NOE-derived distance constraints. Spectra were assigned using 2-dimensional NMR methods at 400 MHz, and internuclear distances were determined from NOESY experiments. Three-bond spin-spin couplings between C alpha H and amide protons, amide exchange rates, and the temperature dependence of amide chemical shifts were also measured. The structure consists largely of loops and turns, with a short region of beta-sheet. The Leu-5 substitution produces a substantial reduction in affinity for trypsin relative to native EETI-II, which contains an Ile at this position. The global structure of the Leu-5 analogue studied here is similar to that reported for native EETI-II (Heitz A, Chiche L, Le-Nguyen D, Castro B, 1989, Biochemistry 28:2392-2398) and to X-ray and NMR structures of the related proteinase inhibitor CMTI-I (Bode W et al., 1989, FEBS Lett 242:285-292; Holak TA et al., 1989a, J Mol Biol 210:649-654; Holak TA, Gondol D, Otlewski J, Wilusz T, 1989b, J Mol Biol 210:635-648; Holak TA, Habazettl J, Oschkinat H, Otlewski J, 1991, J Am Chem Soc 113:3196-3198). The region near the scissile bond is the most disordered part of the structure, based on geometric superimposition of 40 calculated structures. This disorder most likely reflects additional motion being present in this region relative to the rest of the protein. This motional disorder is increased in the Leu-5 analogue relative to the native form and may be responsible for its reduced trypsin binding. A second form of the protein synthesized with all (D) amino acids was also studied by NMR and found to have a spectrum identical with that of the (L) form. This is consistent with the (D) form being a mirror image of the (L) form and not distinguishable by NMR in an achiral solvent (i.e., H2O). The (D) form has no activity against trypsin, as would be expected for a mirror-image form.
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{{ABSTRACT_PUBMED_8003965}}
==About this Structure==
==About this Structure==
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2LET is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Ecballium_elaterium Ecballium elaterium]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2LET OCA].
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2LET is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Ecballium_elaterium Ecballium elaterium]. Full experimental information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2LET OCA].
==Reference==
==Reference==
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[[Category: Kent, S B.H.]]
[[Category: Kent, S B.H.]]
[[Category: Nielsen, K J.]]
[[Category: Nielsen, K J.]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun May 4 09:30:05 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Jul 29 12:03:44 2008''

Revision as of 09:03, 29 July 2008

Image:2let.png

Template:STRUCTURE 2let

AN 1H NMR DETERMINATION OF THE THREE DIMENSIONAL STRUCTURES OF MIRROR IMAGE FORMS OF A LEU-5 VARIANT OF THE TRYPSIN INHIBITOR ECBALLIUM ELATERIUM (EETI-II)

Template:ABSTRACT PUBMED 8003965

About this Structure

2LET is a Single protein structure of sequence from Ecballium elaterium. Full experimental information is available from OCA.

Reference

An 1H NMR determination of the three-dimensional structures of mirror-image forms of a Leu-5 variant of the trypsin inhibitor from Ecballium elaterium (EETI-II)., Nielsen KJ, Alewood D, Andrews J, Kent SB, Craik DJ, Protein Sci. 1994 Feb;3(2):291-302. PMID:8003965

Page seeded by OCA on Tue Jul 29 12:03:44 2008

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