Purification and structural characterization of the CD11b/CD18 integrin α subunit I domain reveals a folded conformation in solution

Fairbanks, Michael B.; Pollock, John R.; Prairie, Mark D.; Scahill, Terrence A.; Baczynskyj, Lubo; Heinrikson, Robert L.; Stockman, Brian J.

doi:10.1016/0014-5793(95)00747-w

Cited by 8 publications

(6 citation statements)

References 31 publications

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“…1A). is very similar to those of other members of the vWA-domain family [14,15] inasmuch as it is also characterized by a large negative ellipticity between 210 and 225 nm and a small trough between these wavelengths.…”

Section: Expression and Characterization Of The Second Vonsupporting

confidence: 61%

The second von Willebrand type A domain of cochlin has high affinity for type I, type II and type IV collagens

2008

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Cochlin is colocalized with type II collagen in the extracellular matrix of cochlea and has been suggested to interact with this collagen. Here we show that the second von Willebrand type A domain of cochlin has affinity for type II collagen, as well as type I and type IV collagens whereas the LCCL-domain of cochlin has no affinity for these proteins. The implications of these findings for the mechanism whereby cochlin mutations cause the dominant negative DFNA9-type hearing loss are discussed.

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Section: Expression and Characterization Of The Second Vonsupporting

confidence: 61%

The second von Willebrand type A domain of cochlin has high affinity for type I, type II and type IV collagens

2008

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“…Several recent publications identified the structure for parts of other integrin subunits by CD spectroscopy e.g. the domain I of CD11b/CD18 leukocyte integrin [35] or the biological active part of α3β1 integrin [36] and even for the cytoplasmic tail and the transmembrane domain of αIIbβ3 [37]. Additionally, MDS studies as well as crystal structures have shown the dynamics of integrin motion [38, 39].…”

Section: Discussionmentioning

confidence: 99%

Drug-induced activation of integrin alpha IIb beta 3 leads to minor localized structural changes

et al. 2019

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Integrins are transmembrane proteins involved in hemostasis, wound healing, immunity and cancer. In response to intracellular signals and ligand binding, integrins adopt different conformations: the bent (resting) form; the intermediate extended form; and the ligand-occupied active form. An integrin undergoing such conformational dynamics is the heterodimeric platelet receptor αIIbβ3. Although the dramatic rearrangement of the overall structure of αIIbβ3 during the activation process is potentially related to changes in the protein secondary structure, this has not been investigated so far in a membrane environment. Here we examine the Mn 2+ - and drug-induced activation of αIIbβ3 and the impact on the structure of this protein reconstituted into liposomes. By quartz crystal microbalance with dissipation monitoring and activation assays we show that Mn 2+ induces binding of the conformation-specific antibody PAC-1, which only recognizes the extended, active integrin. Circular dichroism spectroscopy reveals, however, that Mn 2+ -treatment does not induce major secondary structural changes of αIIbβ3. Similarly, we found that treatment with clinically relevant drugs (e.g. quinine) led to the activation of αIIbβ3 without significant changes in protein secondary structure. Molecular dynamics simulation studies revealed minor local changes in the beta-sheet probability of several extracellular domains of the integrin. Our experimental setup represents a new approach to study transmembrane proteins, especially integrins, in a membrane environment and opens a new way for testing drug binding to integrins under clinically relevant conditions.

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“…The ␤ 3 MIDAS domain is structurally analogous to integrin ␣ subunit I domains from ␣ L and ␣ M , which can be expressed as isolated entities (15)(16)(17), so it is not surprising that it too can be expressed as an isolated domain, as we describe below. However, our parallel objective was to identify a short segment of the ␣ IIb subunit that is also necessary for heterodimer assem- bly.…”

Section: Resultsmentioning

confidence: 99%

Molecular Requirements for Assembly and Function of a Minimized Human Integrin αIIbβ3

McKay

Annis

Honda³

et al. 1996

Journal of Biological Chemistry

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Integrin subunit compatibility within and between species plays a major role in heterodimer assembly and ligand specificity. As an example, human ␣ IIb pairs only with human ␤ 3 and does not assemble a heterodimer with ␤ 3 from other species. We use interspecies subunit chimeras to identify molecular requirements for subunit compatibility and show that species-restricted heterodimer assembly depends on a unique hexapeptide VGSDNH in an extended loop of the hypothetical human ␤ 3 MIDAS domain. This allows us to express ␣ IIb (1-233) and ␤ 3 (111-318) as a soluble, mini-integrin that retains RGD-dependent ligand recognition. Thus, in the case of one integrin, ␣ IIb ␤ 3 , the molecular requirements for integrin subunit compatibility and ligand recognition are intimately related.

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Purification and structural characterization of the CD11b/CD18 integrin α subunit I domain reveals a folded conformation in solution

Cited by 8 publications

References 31 publications

The second von Willebrand type A domain of cochlin has high affinity for type I, type II and type IV collagens

The second von Willebrand type A domain of cochlin has high affinity for type I, type II and type IV collagens

Drug-induced activation of integrin alpha IIb beta 3 leads to minor localized structural changes

Molecular Requirements for Assembly and Function of a Minimized Human Integrin αIIbβ3

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