Purification, crystallization and preliminary X-ray investigation of the complex of human vitamin D binding protein and rabbit muscle actin

Bogaerts, Ilse; Verboven, Christel; Rabijns, A.; Waelkens, Etienne; Baelen, Hugo Van; Ranter, Camiel De

doi:10.1107/s090744490100350x

Cited by 6 publications

(7 citation statements)

References 23 publications

(21 reference statements)

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“…Actin and DBP mixed at a 1.2:1.0 molar ratio and purified on a Superose 12 HR column (running buffer: 100 mM Tris, pH 7.6͞0.6 mM ATP͞0.5 mM ␤-mercaptoethanol͞0.2 mM CaCl 2 ) were concentrated to Ϸ20 mg⅐ml Ϫ1 and crystallized at 4°C in 11% polyethylene glycol 12,000͞200 mM magnesium acetate͞100 mM sodium cacodylate pH 6.6͞20% glycerol. Although these conditions are similar to those of Bogaerts et al (8), our crystals belong to a different space group (P2 1 2 1 2 1 ).…”

Section: Methodsmentioning

confidence: 63%

Crystal structures of the vitamin D-binding protein and its complex with actin: Structural basis of the actin-scavenger system

Otterbein

Cosio

Graceffa

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

149

121

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Actin is the most abundant protein in eukaryotic cells, but its release from cells into blood vessels can be lethal, being associated with clinical situations including hepatic necrosis and septic shock. A homeostatic mechanism, termed the actin-scavenger system, is responsible for the depolymerization and removal of actin from the circulation. During the first phase of this mechanism, gelsolin severs the actin filaments. In the second phase, the vitamin Dbinding protein (DBP) traps the actin monomers, which accelerates their clearance. We have determined the crystal structures of DBP by itself and complexed with actin to 2.1 Å resolution. Similar to its homologue serum albumin, DBP consists of three related domains. Yet, in DBP a strikingly different organization of the domains gives rise to a large actin-binding cavity. After complex formation the three domains of DBP move slightly to ''clamp'' onto actin subdomain 3 and to a lesser extent subdomain 1. Contacts between actin and DBP throughout their extensive 3,454-Å 2 intermolecular interface involve a mixture of hydrophobic, electrostatic, and solventmediated interactions. The area of actin covered by DBP within the complex approximately equals the sum of those covered by gelsolin and profilin. Moreover, certain interactions of DBP with actin mirror those observed in the actin-gelsolin complex, which may explain how DBP can compete effectively with gelsolin for actin binding. Formation of the strong actin-DBP complex proceeds with limited conformational changes to both proteins, demonstrating how DBP has evolved to become an effective actin-scavenger protein.

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Section: Methodsmentioning

confidence: 63%

Crystal structures of the vitamin D-binding protein and its complex with actin: Structural basis of the actin-scavenger system

Otterbein

Cosio

Graceffa

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

149

121

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“…DBP is positioned on chromosome 4 in proximity to chemokines, such as interleukin 8, and is thought to participate in redox-regulated inflammatory responses (Yamamoto and Naraparaju, 1996). Albumin and DBP contain cysteine residues that predict a characteristic pattern of disulfide bridges and homologous protein folding (Bogaerts et al, 2001). DBP also binds globular actin with high affinity and inhibits actin polymerization by sequestering monomeric G-actin, thereby limiting construction of "signaling roadways" (McLeod et al, 1989).…”

Section: Discussionmentioning

confidence: 99%

Identification of Albumin Precursor Protein, Phi AP3, and α-Smooth Muscle Actin as Novel Components of Redox Sensing Machinery in Vascular Smooth Muscle Cells

et al. 2002

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Aerobic organisms are continually subjected to environmental stressors that compromise redox homeostasis and induce cellular injury. In vascular smooth muscle cells (vSMCs), the activation/repression of redox-regulated genes after environmental stress often involves protein binding to cis-acting antioxidant response elements (AREs). The present study was conducted to identify proteins that participate in redox-regulated protein binding to human c-Ha-ras and mouse glutathione S-transferase A1 AREs in vSMCs after oxidant injury. Challenge of vSMCs with 0.3 or 3 M hydrogen peroxide, 3-methylcholanthrene, benzo[a]pyrene-7,8-diol, 3-hydroxy benzo[a]pyrene, and benzo[a]pyrene-3,6-quinone induced concentration-related increases in ARE protein binding. The profiles of ARE complex assembly were comparable, but exhibited chemical specificity. Pretreatment with 0.5 mM N-acetylcysteine inhibited activation of ARE protein binding in hydrogen peroxidetreated cells. Preparative electrophoretic mobility shift assays coupled to Western analysis identified NF-E2-related proteins 1 and 2 and JunD in complexes assembled on AREs. Polyethylenimine affinity and sequence-specific serial immobilized DNA affinity chromatography followed by N-terminal sequencing identified albumin precursor protein, phi AP3, and ␣-smooth muscle actin as members of the ARE signaling pathway. Sequence analysis of albumin protein revealed homology to the redox-regulated transcription factors Bach1 and 2, as well as cytoskeletal and molecular motor proteins. These results implicate albumin precursor protein, phi AP3, and ␣-smooth muscle actin as participants in redox sensing in vSMCs, and suggest that protein complex assembly involves interactions between leucine zipper and zinc finger transcription factors with cytoskeletal proteins.

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“…The crystal T A Asp Lys structure of actin-bound DBP has also been solved. [60][61][62] From these studies, actin subdomains one, and three were determined to bind along a groove comprised of surfaces from all three domains of DBP. This large surface area of contact is greater than other actin binding proteins like gelsolin, profilin and DNase I.…”

Section: Molecular Biology and Biochemistry Of Dbpmentioning

confidence: 99%

“…The third domain is truncated at the C‐terminal end containing only four α‐helices. The crystal structure of actin‐bound DBP has also been solved . From these studies, actin subdomains one, and three were determined to bind along a groove comprised of surfaces from all three domains of DBP.…”

Section: Introductionmentioning

confidence: 99%

New perspectives on the vitamin D binding protein

Chun

2012

Cell Biochemistry & Function

221

150

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The serum vitamin D binding protein (DBP), also known as GC-globulin, is a multifunctional protein known for its role in the transport of vitamin D metabolites. DBP also binds fatty acids and actin monomers, preventing their polymerization that could be detrimental in the circulatory system. DBP may have immune functions independent of its role as a transporter of vitamin D. Because of the abundance of DBP, many aspects of its basic biochemistry were quickly established. Other features of vitamin D action, particularly transcriptional mechanisms of regulation, received greater focus and early interest in DBP centred on its value as a tool for population genetics because of its intriguing genetic variations. Nonetheless, knowledge of DBP mechanisms in physiology was obtained, and functions beyond vitamin D ligand binding were identified. With the recent increased attention regarding the benefits of vitamin D (bone health and immunological regulation), there has been a resurgence of interest in DBP. Because DBP is the primary transporter of vitamin D, it has a role in maintaining the total levels of vitamin D for the organism and in regulating the amounts of free (unbound) vitamin D available for specific tissues and cell types to utilize. This review will describe the findings on the basic biochemistry and molecular biology of DBP, the studies that elucidated its biological functions and highlight these results in light of the current renewed interest in vitamin D and human health, as well as the debate over what constitutes sufficient levels of vitamin D.

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Purification, crystallization and preliminary X-ray investigation of the complex of human vitamin D binding protein and rabbit muscle actin

Cited by 6 publications

References 23 publications

Crystal structures of the vitamin D-binding protein and its complex with actin: Structural basis of the actin-scavenger system

Crystal structures of the vitamin D-binding protein and its complex with actin: Structural basis of the actin-scavenger system

Identification of Albumin Precursor Protein, Phi AP3, and α-Smooth Muscle Actin as Novel Components of Redox Sensing Machinery in Vascular Smooth Muscle Cells

New perspectives on the vitamin D binding protein

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