Universität zu Köln

Abraham, Elena T., Oecal, Sinan, Morgelin, Matthias, Schmid, Philipp W. N., Buchner, Johannes ORCID: 0000-0003-1282-7737, Baumann, Ulrich and Gebauer, Jan M. (2021). Collagen's primary structure determines collagen:HSP47 complex stoichiometry. J. Biol. Chem., 297 (6). AMSTERDAM: ELSEVIER. ISSN 1083-351X

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Abstract

Collagens play important roles in development and homeostasis in most higher organisms. In order to function, collagens require the specific chaperone HSP47 for proper folding and secretion. HSP47 is known to bind to the collagen triple helix, but the exact positions and numbers of binding sites are not clear. Here, we employed a collagen II peptide library to characterize high-affinity binding sites for HSP47. We show that many previously predicted binding sites have very low affinities due to the presence of a negatively charged amino acid in the binding motif. In contrast, large hydrophobic amino acids such as phenylalanine at certain positions in the collagen sequence increase binding strength. For further characterization, we determined two crystal structures of HSP47 bound to peptides containing phenylalanine or leucine. These structures deviate significantly from previously published ones in which different collagen sequences were used. They reveal local conformational rearrangements of HSP47 at the binding site to accommodate the large hydrophobic side chain from the middle strand of the collagen triple helix and, most surprisingly, possess an altered binding stoichiometry in the form of a 1:1 complex. This altered stoichiometry is explained by steric collisions with the second HSP47 molecule present in all structures determined thus far caused by the newly introduced large hydrophobic residue placed on the trailing strand. This exemplifies the importance of considering all three sites of homotrimeric collagen as independent interaction surfaces and may provide insight into the formation of higher oligomeric complexes at promiscuous collagen-binding sites.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Abraham, Elena T. UNSPECIFIED UNSPECIFIED UNSPECIFIED Oecal, Sinan UNSPECIFIED UNSPECIFIED UNSPECIFIED Morgelin, Matthias UNSPECIFIED UNSPECIFIED UNSPECIFIED Schmid, Philipp W. N. UNSPECIFIED UNSPECIFIED UNSPECIFIED Buchner, Johannes UNSPECIFIED orcid.org/0000-0003-1282-7737 UNSPECIFIED Baumann, Ulrich UNSPECIFIED UNSPECIFIED UNSPECIFIED Gebauer, Jan M. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-581838
DOI:	10.1016/j.jbc.2021.101169
Journal or Publication Title:	J. Biol. Chem.
Volume:	297
Number:	6
Date:	2021
Publisher:	ELSEVIER
Place of Publication:	AMSTERDAM
ISSN:	1083-351X
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language CHAPERONE HSP47; BINDING-SITES; TRIPLE-HELIX; RECOGNITION; PROTEIN Multiple languages Biochemistry & Molecular Biology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/58183