Structural elucidation of full-length nidogen and the laminin-nidogen complex in solution.
Nidogen-1 is a key basement membrane protein that is required for many biological activities. It is one of the central elements in organizing basal laminae including those in the skin, muscle, and the nervous system. The self-assembling extracellular matrix that also incorporates fibulins, fibronectin and integrins is clamped together by networks formed between nidogen, perlecan, laminin and collagen IV. To date, the full-length version of nidogen-1 has not been studied in detail in terms of its solution conformation and shape because of its susceptibility to proteolysis. In the current study, we have expressed and purified full-length nidogen-1 and have investigated its solution behavior using size-exclusion chromatography (SEC), dynamic light scattering (DLS) and small angle X-ray scattering (SAXS). The ab initio shape reconstruction of the complex between nidogen-1 and the laminin γ-1 short arm confirms that the interaction is mediated solely by the C-terminal domains: the rest of the domains of both proteins do not participate in complex formation.
Department of Chemistry, University of Manitoba, 144 Dysart Road, Winnipeg, Manitoba RT3 2N2, Canada.
Trushar R
TR
Patel
Institute for Dental Research and Oral Musculoskeletal Biology, Center for Molecular Medicine Cologne, Center for Biochemistry, Medical Faculty, University of Cologne, D-50931 Cologne, Germany.
Claudia
C
Bernards
Department of Chemistry, University of Manitoba, 144 Dysart Road, Winnipeg, Manitoba RT3 2N2, Canada.
Markus
M
Meier
Department of Chemistry, University of Manitoba, 144 Dysart Road, Winnipeg, Manitoba RT3 2N2, Canada.
Kevin
K
McEleney
School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, Queensland, 4072, Australia.
Donald J
DJ
Winzor
Institute for Dental Research and Oral Musculoskeletal Biology, Center for Molecular Medicine Cologne, Center for Biochemistry, Medical Faculty, University of Cologne, D-50931 Cologne, Germany.
Manuel
M
Koch
Department of Chemistry, University of Manitoba, 144 Dysart Road, Winnipeg, Manitoba RT3 2N2, Canada; Microbiology, University of Manitoba, 144 Dysart Road, Winnipeg, Manitoba RT3 2N2, Canada; Biochemistry and Medical Genetics, University of Manitoba, 144 Dysart Road, Winnipeg, Manitoba RT3 2N2, Canada. Electronic address: jorg.stetefeld@ad.manitoba.ca.
Jörg
J
Stetefeld
D000818:Animals; D007797:Laminin / Q000737:chemistry*; D008562:Membrane Glycoproteins / Q000737:chemistry*; D051379:Mice; D008958:Models, Molecular; D010316:Particle Size; D054730:Protein Interaction Domains and Motifs; D020836:Protein Structure, Quaternary; D053838:Scattering, Small Angle; D012996:Solutions; D014961:X-Ray Diffraction
D007797:Laminin; D008562:Membrane Glycoproteins; D012996:Solutions; C121346:laminin 1; C029967:nidogen
Dynamic light scattering; Extracellular matrix proteins; Hydrodynamics; Laminin; Nidogen-1; Small angle X-ray scattering