Disease relevance of HSAB

• Either ethyldimethylisopropyl-aminocarbodiimide or hydroxysuccinimidyl-p-azidobenzoate causes highly specific crosslinking of 125I-NGF to a similar receptor species on rat pheochromocytoma PC12 cells and on human melanoma A875 cells [1].
• This band was formed only in the presence of the HSAB cross-linker when virions were bound to the cells [2].
• We used photoaffinity cross-linking with the heterobifunctional cross-linker N-hydroxysuccinimidyl 4-azidobenzoate (HSAB) to covalently link polyomavirus to a mouse kidney cell surface component [2].
• Clonal PC12 rat pheochromocytoma cells were sequentially incubated with 125I-labeled nerve growth factor and the photoreactive bifunctional agent hydroxysuccinimidyl-p-azidobenzoate [3].

High impact information on HSAB

• In addition, T cell activation by HSAB-coupled beads was specifically inhibited by soluble monomeric HSAB-glycine [4].
• To examine T lymphocyte recognition of foreign antigen, specific responses to the photoreactive antigen N-hydroxysuccinimidyl 4-azidobenzoate (HSAB) were determined by using an HSAB/I-Ad-reactive murine T cell hybridoma [4].
• This stimulation was specific for the phenyl ring composition of HSAB and for HSAB-reactive T cells [4].
• Cross-linking of 125I-nerve growth factor (NGF) to PC12 cells with the photoreactive heterobifunctional agent N-hydroxysuccinimidyl-4-azidobenzoate results in the labeling of two major bands with Mr 158,000 and 100,000 and a minor band with Mr 225,000 as determined by polyacrylamide gel electrophoresis under denaturing and reducing conditions [5].
• Human follicle-stimulating hormone (hFSH) was acylated with N-hydroxysuccinimidyl-4-azidobenzoate (HSAB) and radioiodinated (55 microCi/micrograms) for use as a photoaffinity probe to investigate the subunit structure of the FSH receptor in calf testis [6].

Biological context of HSAB

• In order to determine the binding site of omega to RNA polymerase, we cross-linked omega to RNA polymerase with the hetero-bifunctional cross-linker N-hydroxysuccinimidyl 4-azidobenzoate and analyzed for cross-linked partners using antibodies raised against each of the subunits [7].

Anatomical context of HSAB

• Membranes from adult rabbit superior cervical ganglia, cross-linked to membrane-bound 125I-labeled nerve growth factor (NGF) by the photoreactive agent hydroxysuccinimidyl-p-azidobenzoate, were found to contain two labeled components with apparent Mr = 143,000 and Mr = 112,000 [8].
• In order to determine the optimal reaction conditions, pig liver membranes were incubated with 5 nM [8-lysine]vasopressin (LVP) labeled with 125I and then cross-linked with the use of DMS (dimethyl suberimidate), EGS [ethylene glycol bis(succinimidyl succinate)] or HSAB (hydroxysuccinimidyl p-azidobenzoate) at different final concentrations [9].
• [125I]Calmodulin binding to isolated rat adipocyte plasma membranes has been characterized, and the calmodulin binding proteins associated with the membrane have been identified by use of the photoaffinity cross-linker N-hydroxysuccinimidyl 4-azidobenzoate [10].
• TSH receptors from guinea pig thyroid and epididymal fat have been covalently crosslinked to 125I-labelled TSH conjugated to N-hydroxysuccinimidyl-4-azidobenzoate [11].
• Somatostatin receptors in rat corpus striatum and anterior pituitary membranes were covalently bound to the non-reducible somatostatin analog, [125I]CGP 23996, using the crosslinking agent n-hydroxysuccinimidyl-4-azidobenzoate and ultraviolet light [12].

Associations of HSAB with other chemical compounds

• 125I-SS-28-labeled beta membranes were successfully cross-linked using either the cleavable cross-linker dithiobis(succinimidylpropionate) (1 mM) alone or with a heterobifunctional agent, N-hydroxysuccinimidyl-4-azidobenzoate (HSAB) [13].
• Cross-linking studies with disuccinimidyl suberate and N-hydroxysuccinimidyl-4-azidobenzoate using membrane-solubilised, partially purified and CGRP-affinity-purified preparations, show a number of 125I-alpha-CGRP binding subunit(s) of Mr 62,000-68,000 [14].

Gene context of HSAB

• N-Hydroxysuccinimidyl 4-azidobenzoate, a u.v.-sensitive heterobifunctional reagent, was used to synthesize photoreactive derivatives of the vasoactive intestinal peptide (VIP) [15].
• The hepatic glucagon receptor was covalently labeled with [125I-Try10]monoiodoglucagon [( 125I]MIG) by use of the heterobifunctional cross-linker hydroxysuccinimidyl p-azidobenzoate [16].
• The high affinity somatostatin receptors were covalently labeled with [125I] CGP 23996 and the photoactivated crosslinking agent n-hydroxysuccinimidyl-4-azidobenzoate [17].
• Photoaffinity labeling of the PTH/PTHrP receptors using N-hydroxysuccinimidyl-4-azidobenzoate modified [125I]chPTHrP(1-36) revealed the same specifically labeled 90 kDa protein in CHO-E2 and OK cells [18].
• Cross-linking of the PTH/PTHrP receptors with [125I]chPTHrP-(1-36) modified with N-hydroxysuccinimidyl-4-azidobenzoate revealed indistinguishable doublets of 83 and 73 kilodaltons in both BBM and BLM [19].

Analytical, diagnostic and therapeutic context of HSAB

• Porcine thyrotropin (TSH) receptors have been purified by Sepharose-TSH affinity chromatography and crosslinked to a 125I-labelled photoactive derivative (N-hydroxysuccinimidyl 4-azidobenzoate; HSAB) of TSH (125I-HSAB-TSH) [20].
• The V1-ag, d(CH2)5Tyr(Me)Tyr(NH2)AVP was modified by radiomonoiodination of d(CH2)5-Tyr(Me)Tyr(NH2)AVP with the Iodogen technique, and derivatization of d(CH2)5Tyr(Me)Tyr[125I](NH2)-AVP with the photoreactive crosslinker, N-hydroxysuccinimidyl-4-azidobenzoate (HSAB) (each step included HPLC purification) [21].
• After chromatography, identification of proteins as NGFRc was verified both by immunoprecipitation using receptor-specific monoclonal antibodies and by covalent cross-linking to 125I-NGF using N-hydroxysuccinimidyl-4-azidobenzoate [22].

References