1nub

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{{STRUCTURE_1nub| PDB=1nub | SCENE= }}
{{STRUCTURE_1nub| PDB=1nub | SCENE= }}
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'''HELIX C DELETION MUTANT OF BM-40 FS-EC DOMAIN PAIR'''
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===HELIX C DELETION MUTANT OF BM-40 FS-EC DOMAIN PAIR===
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==Overview==
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The extracellular calcium-binding domain (positions 138-286) of the matrix protein BM-40 possesses a binding epitope of moderate affinity for several collagen types. This epitope was predicted to reside in helix alphaA and to be partially masked by helix alphaC. Here we show that deletion of helix alphaC produces a 10-fold increase in collagen affinity similar to that seen after proteolytic cleavage of this helix. The predicted removal of the steric constraint was clearly demonstrated by the crystal structure of the mutant at 2.8 A resolution. This constitutively activated mutant was used to map the collagen-binding site following alanine mutagenesis at 13 positions. Five residues were crucial for binding, R149 and N156 in helix alphaA, and L242, M245 and E246 in a loop region connecting the two EF hands of BM-40. These residues are spatially close and form a flat ring of 15 A diameter which matches the diameter of a triple-helical collagen domain. The mutations showed similar effects on binding to collagens I and IV, indicating nearly identical binding sites on both collagens. Selected mutations in the non-activated mutant DeltaI also reduced collagen binding, consistent with the same location of the epitope but in a more cryptic form in intact BM-40.
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(as it appears on PubMed at http://www.pubmed.gov), where 9501084 is the PubMed ID number.
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{{ABSTRACT_PUBMED_9501084}}
==About this Structure==
==About this Structure==
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[[Category: Glycosylated protein]]
[[Category: Glycosylated protein]]
[[Category: Site-directed mutagenesis]]
[[Category: Site-directed mutagenesis]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 02:59:21 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Jul 28 20:58:36 2008''

Revision as of 17:58, 28 July 2008

Image:1nub.png

Template:STRUCTURE 1nub

HELIX C DELETION MUTANT OF BM-40 FS-EC DOMAIN PAIR

Template:ABSTRACT PUBMED 9501084

About this Structure

1NUB is a Single protein structure of sequence from Homo sapiens. Full crystallographic information is available from OCA.

Reference

Crystal structure and mapping by site-directed mutagenesis of the collagen-binding epitope of an activated form of BM-40/SPARC/osteonectin., Sasaki T, Hohenester E, Gohring W, Timpl R, EMBO J. 1998 Mar 16;17(6):1625-34. PMID:9501084

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