Cross-linking of protein by ω-maleimido alkanoylN-hydroxysuccinimido esters

Partis, Michael D.; Griffiths, David G.; Roberts, Gretel; Beechey, R. B.

doi:10.1007/bf01025358

Cited by 156 publications

(116 citation statements)

References 26 publications

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“…5). This reaction is extremely selective in the pH range 6.5-7.5 47 and only at higher pH values some cross- reactivity with amino groups may be observed. There are a great variety of commercial available reagents that can be used for introducing maleimide groups on an amine-derivatized surface.…”

Section: Immobilization Of Thiol-containing Proteinsmentioning

confidence: 99%

New Developments for the Site-Specific Attachment of Protein to Surfaces

Camarero

2006

Biophys. Rev. Lett.

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Protein immobilization on surfaces is of great importance in numerous applications in biology and biophysics. The key for the success of all these applications relies on the immobilization technique employed to attach the protein to the corresponding surface. Protein immobilization can be based on covalent or noncovalent interaction of the molecule with the surface. Noncovalent interactions include hydrophobic interactions, hydrogen bonding, van der Waals forces, electrostatic forces, or physical adsorption. However, since these interactions are weak, the molecules can get denatured or dislodged, thus causing loss of signal. They also result in random attachment of the protein to the surface. Site-specific covalent attachment of proteins onto surfaces, on the other hand, leads to molecules being arranged in a definite, orderly fashion and uses spacers and linkers to help minimize steric hindrances between the protein surface. This work reviews in detail some of the methods most commonly used as well as the latest developments for the site-specific covalent attachment of protein to solid surfaces.

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Section: Immobilization Of Thiol-containing Proteinsmentioning

confidence: 99%

New Developments for the Site-Specific Attachment of Protein to Surfaces

Camarero

2006

Biophys. Rev. Lett.

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“…The heterogeneity observed upon cross-linking is probably due to the reaction mechanisms of BS 3 and disuccinimidyl glutarate. Their principal targets are primary amines, such as the amino group at the N terminus of the peptide chains and the ⑀-amino group of lysine residues (30). The human matrilin-1 domain has four lysines.…”

Section: Observed (Data Not Shown)mentioning

confidence: 99%

The C-terminal Domain of Matrilin-2 Assembles into a Three-stranded α-Helical Coiled Coil

Pan¹,

Beck²

1998

Journal of Biological Chemistry

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Matrilin-2 is a member of von Willebrand factor A containing extracellular matrix proteins in which the cDNA-derived sequence shows similar domain organization to cartilage matrix protein/matrilin-1, but information on the protein structure is limited. Here we studied the oligomerization potential of a synthetic peptide NH 2 -ENLILFQNVANEEVRKLTQRLEEMTQRMEALE-NRLKYR-COOH corresponding to the C-terminal sequence of mouse matrilin-2. The central portion of this sequence shows a periodicity of hydrophobic residues occupying positions a and d of a heptad pattern (abcdefg) n , which is characteristic for ␣-helical coiled-coil proteins. Circular dichroism spectroscopy revealed a high ␣-helical content, and the shape of the spectra is indicative for a coiled-coil conformation. Chemical cross-linking and size exclusion chromatography suggest a homotrimeric configuration. Thermal denaturation in benign buffer shows a single cooperative transition with ⌬H 0 ‫؍‬ ؊375 kJ/mol. Melting temperatures T m varied from 38 to 51°C within a concentration range of 10 to 85 M, which is about 35°C lower than determined for a peptide corresponding to the C-terminal domain of matrilin-1. The data suggest that despite the low sequence identity within this region, matrilin-2 will form a homotrimer as matrilin-1 does.Matrilins form a subfamily of extracellular matrix proteins containing von Willebrand factor A-like domains. Its prototype member, matrilin-1 (also known as cartilage matrix protein), is specifically localized in some types of the hyaline cartilage (1) where it can interact with aggrecan and collagen type II fibrils (2-4), but it can also form a filamentous network by itself (5). Its primary structure contains two N-terminal sequence segments with similarities to the von Willebrand factor A domain separated by an epidermal growth factor-like domain and a short unique C-terminal domain (6 -9) that was recently found to be responsible for the oligomerization into homotrimers by assembling into a three-stranded ␣-helical coiled coil (4, 10, 11). Meanwhile, cDNA-derived sequences of two different gene products have been established that show a similar domain structure and are consequently named matrilin-2 and matrilin-3 (12-14). Within the matrilin-2 sequence, the two von Willebrand factor domains are separated by 10 epidermal growth factor-like domains, whereas matrilin-3 lacks the second von Willebrand factor domain and contains four epidermal growth factor-like tandem repeats. In contrast to matrilin-1, matrilin-2 is absent from epiphyseal and other cartilages, but is found in high abundance in the limbs, calvaria, uterus, and heart and in lower amounts in skeletal muscle, brain, and skin (12). Expression studies suggest a similar tissue distribution for matrilin-3 as found for matrilin-1. It mainly co-localizes with collagen type II, especially in the periphery of cartilage in sternum, femur, and trachea (13,14).Currently the oligomerization state of the new matrilins is unknown. Preliminary data from SDS-polyacrylamide gel elec...

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“…65 After incubation for 30 min at room temperature, the cross-linker was quenched by addition of 1 M Tris-HCl, pH 7.5, to a final concentration of 20 mM. After quenching, membranes were lysed in RIPA buffer (10 mM HEPES at pH 7.5, 1% NP-40, 0.1% sodium deoxycholate, 0.1% SDS, 0.15 M NaCl, 1 mM EDTA, 10 mg/ml aprotinin) and cleared by centrifugation at 12 0006g.…”

Section: Cross-linkingmentioning

confidence: 99%

Calphostin C-mediated translocation and integration of Bax into mitochondria induces cytochrome c release before mitochondrial dysfunction

et al. 2000

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Calphostin C-mediated apoptosis in glioma cells was reported previously to be associated with down-regulation of Bcl-2 and Bcl-x L . In this study, we report that 100 nM calphostin C also induces translocation and integration of monomeric Bax into mitochondrial membrane, followed by cytochrome c release into cytosol and subsequent decrease of mitochondrial inner membrane potential (DCm) before activation of caspase-3. The integration of monomeric Bax was associated with acquirement of alkali-resistance. The translocated monomeric Bax was partly homodimerized after cytochrome c release and decrease of DCm. The translocation and homodimerization of Bax, cytochrome c release, and decrease of DCm were not blocked by 100 mM z-VAD.fmk, a pan-caspase inhibitor, but the homodimerization of Bax and decrease of DCm were inhibited by 10 mM oligomycin, a mitochondrial F 0 F 1 -ATPase inhibitor. Therefore, it would be assumed that mitochondrial release of cytochrome c results from translocation and integration of Bax and is independent of permeability transition of mitochondria and caspase activation, representing a critical step in calphostin C-induced cell death. Cell Death and Differentiation (2000) 7, 511 ± 520.

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Cross-linking of protein by ω-maleimido alkanoylN-hydroxysuccinimido esters

Cited by 156 publications

References 26 publications

New Developments for the Site-Specific Attachment of Protein to Surfaces

New Developments for the Site-Specific Attachment of Protein to Surfaces

The C-terminal Domain of Matrilin-2 Assembles into a Three-stranded α-Helical Coiled Coil

Calphostin C-mediated translocation and integration of Bax into mitochondria induces cytochrome c release before mitochondrial dysfunction

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