Site-Specific Cross-Linking of Human and Bovine Hemoglobins Differentially Alters Oxygen Binding and Redox Side Reactions Producing Rhombic Heme and Heme Degradation†

Two new types of covalently linked, site-specific immunoprobes have been prepared using metal cluster labels, and used to stain components of cells. Combined fluorescein and 1.4 nm “Nanogold” labels were prepared by using the fluorescein-conjugated tris (aryl) phosphine ligand and the amino-substituted ligand in the synthesis of the Nanogold cluster. This cluster label was activated by reaction with a 60-fold excess of (sulfo-Succinimidyl-4-N-maleiniido-cyclohexane-l-carboxylate (sulfo-SMCC) at pH 7.5, separated from excess cross-linking reagent by gel filtration, and mixed in ten-fold excess with Goat Fab’ fragments against mouse IgG (obtained by reduction of F(ab’)2 fragments with 50 mM mercaptoethylamine hydrochloride). Labeled Fab’ fragments were isolated by gel filtration HPLC (Superose-12, Pharmacia). A combined Nanogold and Texas Red label was also prepared, using a Nanogold cluster derivatized with both and its protected analog: the cluster was reacted with an eight-fold excess of Texas Red sulfonyl chloride at pH 9.0, separated from excess Texas Red by gel filtration, then deprotected with HC1 in methanol to yield the amino-substituted label.

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Site-specific cross-linking of TBP in vivo and in vitro reveals a direct functional interaction with the SAGA subunit Spt3

Genes & Development ◽

10.1101/gad.1724408 ◽

2008 ◽

Vol 22 (21) ◽

pp. 2994-3006 ◽

Cited By ~ 68

Author(s):

N. Mohibullah ◽

S. Hahn

Keyword(s):

Cross Linking ◽

Functional Interaction ◽

Site Specific

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Alteration of the substrate specificity of human lysozyme by site-specific intermolecular cross-linking

FEBS Letters ◽

10.1016/0014-5793(94)01168-0 ◽

1994 ◽

Vol 355 (3) ◽

pp. 271-274 ◽

Cited By ~ 3

Author(s):

Michiro Muraki ◽

Yoshifumi Jigami ◽

Kazuaki Harata

Keyword(s):

Substrate Specificity ◽

Cross Linking ◽

Human Lysozyme ◽

Site Specific

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Folic Acid-Targeted Disulfide-Based Cross-Linking Micelle for Enhanced Drug Encapsulation Stability and Site-Specific Drug Delivery Against Tumors [Corrigendum]

International Journal of Nanomedicine ◽

10.2147/ijn.s347786 ◽

2021 ◽

Vol Volume 16 ◽

pp. 7683-7684

Author(s):

Yumin Zhang ◽

Junhui Zhou ◽

Cuihong Yang ◽

Weiwei Wang ◽

Liping Chu ◽

...

Keyword(s):

Drug Delivery ◽

Folic Acid ◽

Cross Linking ◽

Specific Drug ◽

Drug Encapsulation ◽

Site Specific

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Site-specific cross-linking of proteins to DNA via a new bioorthogonal approach employing oxime ligation

Chemical Communications ◽

10.1039/c8cc01300d ◽

2018 ◽

Vol 54 (49) ◽

pp. 6296-6299 ◽

Cited By ~ 2

Author(s):

Suresh S. Pujari ◽

Yi Zhang ◽

Shaofei Ji ◽

Mark D. Distefano ◽

Natalia Y. Tretyakova

Keyword(s):

Cross Linking ◽

Cross Link ◽

Site Specific ◽

Link Formation

Model site-specific DNA–protein cross-link formation by bioorthogonal oxime ligation.

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Furan warheads for covalent trapping of weak protein-protein interactions: cross-linking of thymosin β4 to actin

Chemical Communications ◽

10.1039/d1cc01731d ◽

2021 ◽

Author(s):

Laia Miret Casals ◽

Willem Vannecke ◽

Kurt Hoogewijs ◽

Gianluca Arauz ◽

Marina Gay ◽

...

Keyword(s):

Protein Interaction ◽

Protein Interactions ◽

3D Structure ◽

Cross Linking ◽

Thymosin Β4 ◽

Protein Protein Interactions ◽

Site Specific ◽

Protein Protein Interaction ◽

Covalent Trapping

We describe furan as a triggerable ‘warhead’ for site-specific cross-linking using the actin and thymosin β4 (Tβ4)-complex as model of a weak and dynamic protein-protein interaction with known 3D structure...

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