Identifying the SPARC binding sites on collagen I and procollagen I by atomic force microscopy

Hua Wang, Andrzej Fertala, Buddy D. Ratner, E. Helene Sage, Shaoyi Jiang

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

SPARC (secreted protein acidic and rich in cysteine) is a matricellular protein associated with the extracellular matrix (ECM). It has been found that the production of collagen I is a requisite for the association of SPARC with ECM, and studies with SPARC-null mice indicate that SPARC plays a role in modifying the structure of collagen fibers. It is not known, however, whether SPARC interacts with the collagen I precursor, procollagen I. In this study, the binding of SPARC to collagen I and procollagen I was verified by surface plasmon resonance. The SPARC-binding sites on collagen I and procollagen I were identified by directly visualizing their complexes using tapping-mode atomic force microscopy (TM-AFM). The characteristic chain end feature in collagen I is not readily detected by AFM, so unambiguous location of the binding sites relative to the C- or N-termini is difficult. In contrast, procollagen I, with its large globular C-propeptide, permits easy identification of the C-terminus. Histograms were constructed and compared based on the distances of the bound SPARC to the C-terminus of procollagen I and to the closest end of collagen I. There is a broad distribution of SPARC binding sites on procollagen I with the most preferred binding region located ∼1/3 from the C-terminus. Characterization of the SPARC-binding sites on collagen I and procollagen I provides useful information for further understanding of the functional implications of their interactions.

Original languageEnglish (US)
Pages (from-to)6765-6771
Number of pages7
JournalAnalytical Chemistry
Volume77
Issue number21
DOIs
StatePublished - Nov 1 2005
Externally publishedYes

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Procollagen
Cysteine
Atomic force microscopy
Collagen
Binding Sites
Proteins
Surface plasmon resonance

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Identifying the SPARC binding sites on collagen I and procollagen I by atomic force microscopy. / Wang, Hua; Fertala, Andrzej; Ratner, Buddy D.; Sage, E. Helene; Jiang, Shaoyi.

In: Analytical Chemistry, Vol. 77, No. 21, 01.11.2005, p. 6765-6771.

Research output: Contribution to journalArticle

Wang, Hua ; Fertala, Andrzej ; Ratner, Buddy D. ; Sage, E. Helene ; Jiang, Shaoyi. / Identifying the SPARC binding sites on collagen I and procollagen I by atomic force microscopy. In: Analytical Chemistry. 2005 ; Vol. 77, No. 21. pp. 6765-6771.
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