Cyanogen bromide cleaves Fc fragments of pooled human IgG at both methionine and tryptophan residues

Boulware, Dennis W.; Goldsworthy, Patrick D.; Nardella, Francis A.; Mannik, Mart

doi:10.1016/0161-5890(85)90052-5

Cited by 5 publications

(4 citation statements)

References 14 publications

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“…This region is well conserved from bacteria to mammals and the residue at this position was already identified as tryptophan (11, see Supplemental materials). As it was reported that tryptophan residue was oxidized by cyanogen bromide (14), a similar reaction should occur in this case. Furthermore, the product of cycle 9 was not detected at the expected retention time of PTH‐Tyr derivative on HPLC (see Supplemental materials).…”

Section: Resultssupporting

confidence: 72%

Photochemical Nucleophile Mapping: Identification of Tyr311 Within the Catalytic Domain of Rabbit Muscle Glyceraldehyde‐3‐phosphate Dehydrogenase^†

Hatanaka

Kaneda

Tomohiro

2007

Photochem & Photobiology

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Photochemical mapping of nucleophiles in close proximity to the active site Cys149 of rabbit glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was demonstrated based on the nucleophilic aromatic photosubstitution reaction using two regioisomers of alkoxy-fluoro-nitro-substituted benzenes. Two photophores were covalently attached to the active site SH group of GAPDH and the protein was subjected to photolysis then to the cyanogen bromide cleavage reaction. The advantage of this method is the capability to chase labeled products by monitoring absorption at 380 nm because of the chromogenic property of photophore. HPLC separation identified a large labeled peptide fragment that was further digested by V8 protease for Edman sequence analysis. From the recent X-ray crystallography of rabbit GAPDH, Tyr311, His176, Ser238 and Lys183 are closely located to catalytic Cys149. Among these nucleophiles, Tyr311 was preferentially labeled with 2-fluoro-4-nitrophenoxy photophore and no label was identified with the isomeric 4-fluoro-2-nitrophenoxy photophore. The result clearly reflects the distance between Cys149 and nucleophiles to distinguish the nearest Tyr311. As photophores show great reactivity even with water under neutral conditions, the distance between nucleophiles and photophores is important for photoinduced nucleophilic aromatic substitution. The method will provide a useful technique to survey nucleophiles within the catalytic domain.

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Section: Resultssupporting

confidence: 72%

Photochemical Nucleophile Mapping: Identification of Tyr311 Within the Catalytic Domain of Rabbit Muscle Glyceraldehyde‐3‐phosphate Dehydrogenase^†

Hatanaka

Kaneda

Tomohiro

2007

Photochem & Photobiology

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“…Four out of five CNBr fragments were generated by cleavages at the Cterminus side of tryptophan but not at methionine residues. The reason for such frequent cleavage at tryptophan is unclear, although cleavage at tryptophan has been reported with other proteins [33]. Multiple peaks were observed in each cycle of the N-terminal amino acid sequence determination of chitinases C1 and C2 as described above.…”

Section: Cloning and Nucleotide Sequence Of The Gene Encoding Chitinamentioning

confidence: 88%

The third chitinase gene (chiC) of Serratia marcescens 2170 and the relationship of its product to other bacterial chitinases

Suzuki

Taiyoji

Sato

et al. 1999

Biochemical Journal

142

123

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The third chitinase gene (chiC) of Serratia marcescens 2170, specifying chitinases C1 and C2, was identified. Chitinase C1 lacks a signal sequence and consists of a catalytic domain belonging to glycoside hydrolase family 18, a fibronectin type III-like domain (Fn3 domain) and a C-terminal chitin-binding domain (ChBD). Chitinase C2 corresponds to the catalytic domain of C1 and is probably generated by proteolytic removal of the Fn3 and ChBDs. The loss of the C-terminal portion reduced the hydrolytic activity towards powdered chitin and regenerated chitin, but not towards colloidal chitin and glycol chitin, illustrating the importance of the ChBD for the efficient hydrolysis of crystalline chitin. Phylogenetic analysis showed that bacterial family 18 chitinases can be clustered in three subfamilies which have diverged at an early stage of bacterial chitinase evolution. Ser. marcescens chitinase C1 is found in one subfamily, whereas chitinases A and B of the same bacterium belong to another subfamily. Chitinase C1 is the only Ser. marcescens chitinase that has an Fn3 domain. The presence of multiple, divergent, chitinases in a single chitinolytic bacterium is perhaps necessary for efficient synergistic degradation of chitin.

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“…In the CFC region, there are three Lys residues that might be cleaved by endoproteinase Lys‐C and two Trp residues that could be oxidized during CNBr treatment [23]. Additional cleavage can take place on the C‐terminal side of oxidized Trp [24]. The observed protonated mass (MH + ) of the major component in the pooled fractions containing the CFC domain was 4702.4 Da (Fig.…”

Section: Determination Of Disulfide Linkages In the Cfc Domainmentioning

confidence: 99%

The CRIPTO/FRL‐1/CRYPTIC (CFC) domain of human Cripto

Foley

Vlijmen

Boynton

et al. 2003

European Journal of Biochemistry

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The disulfide structure of the CRIPTO/FRL-1/CRYPTIC (CFC) domain of human Cripto protein was determined by a combination of enzymatic and chemical fragmentation, followed by chromatographic separation of the fragments, and characterization by mass spectrometry and N-terminal sequencing. These studies showed that Cys115 forms a disulfide bond with Cys133, Cys128 with Cys149, and Cys131 with Cys140. Protein database searching and molecular modeling revealed that the pattern of disulfide linkages in the CFC domain of Cripto is the same as that in PARS intercerebralis major Peptide C (PMP-C), a serine protease inhibitor, and that the EGF-CFC domains of Cripto are predicted to be structurally homologous to the EGF-VWFC domains of the C-terminal extracellular portions of Jagged 1 and Jagged 2. Biochemical studies of the interactions of ALK4 with the CFC domain of Cripto by fluorescence-activated cell sorter analysis indicate that the CFC domain binds to ALK4 independent of the EGF domain. A molecular model of the CFC domain of Cripto was constructed based on the nuclear magnetic resonance structure of PMP-C. This model reveals a hydrophobic patch in the domain opposite to the presumed ALK4 binding site. This hydrophobic patch may be functionally important for the formation of intra or intermolecular complexes.

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Cyanogen bromide cleaves Fc fragments of pooled human IgG at both methionine and tryptophan residues

Cited by 5 publications

References 14 publications

Photochemical Nucleophile Mapping: Identification of Tyr311 Within the Catalytic Domain of Rabbit Muscle Glyceraldehyde‐3‐phosphate Dehydrogenase^†

Photochemical Nucleophile Mapping: Identification of Tyr311 Within the Catalytic Domain of Rabbit Muscle Glyceraldehyde‐3‐phosphate Dehydrogenase^†

The third chitinase gene (chiC) of Serratia marcescens 2170 and the relationship of its product to other bacterial chitinases

The CRIPTO/FRL‐1/CRYPTIC (CFC) domain of human Cripto

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Cyanogen bromide cleaves Fc fragments of pooled human IgG at both methionine and tryptophan residues

Cited by 5 publications

References 14 publications

Photochemical Nucleophile Mapping: Identification of Tyr311 Within the Catalytic Domain of Rabbit Muscle Glyceraldehyde‐3‐phosphate Dehydrogenase†

Photochemical Nucleophile Mapping: Identification of Tyr311 Within the Catalytic Domain of Rabbit Muscle Glyceraldehyde‐3‐phosphate Dehydrogenase†

The third chitinase gene (chiC) of Serratia marcescens 2170 and the relationship of its product to other bacterial chitinases

The CRIPTO/FRL‐1/CRYPTIC (CFC) domain of human Cripto

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Photochemical Nucleophile Mapping: Identification of Tyr311 Within the Catalytic Domain of Rabbit Muscle Glyceraldehyde‐3‐phosphate Dehydrogenase^†

Photochemical Nucleophile Mapping: Identification of Tyr311 Within the Catalytic Domain of Rabbit Muscle Glyceraldehyde‐3‐phosphate Dehydrogenase^†