Carboxymethylation of Cysteine Using Iodoacetamide/ Iodoacetic Acid

Aitken, Alastair; Learmonth, Michèle

doi:10.1385/1-59259-169-8:455

Cited by 20 publications

(17 citation statements)

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“…Free SH groups were identified using 5,5Ј-dithiobis(2-nitrobenzoic acid) (DTNB; Sigma, St. Louis, Mo.) in native and denaturing buffers (1). Sodium phosphate (0.1 M; pH 8.0) was used as the native buffer.…”

Section: Methodsmentioning

confidence: 99%

Properties and Mutation Analysis of the CelK Cellulose-Binding Domain from the Clostridium thermocellum Cellulosome

Kataeva

Seidel

et al. 2001

J Bacteriol

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Section: Methodsmentioning

confidence: 99%

Properties and Mutation Analysis of the CelK Cellulose-Binding Domain from the Clostridium thermocellum Cellulosome

Kataeva

Seidel

et al. 2001

J Bacteriol

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“…Cysteine was also carboxymethylated following the method of Aitken and Learmonth [25], using β-mercaptoethanol and iodoacetamide. Arginine was modified according to the protocol of Tawfik [26], using p-hydroxyphenylglyoxal.…”

Section: Chemical Modification Of Amino-acid Residuesmentioning

confidence: 99%

Structural analysis and binding properties of isoforms of tarin, the GNA-related lectin from Colocasia esculenta

Pereira¹,

Winter²,

Verícimo³

et al. 2015

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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The lectins, a class of proteins that occur widely in animals, plants, fungi, lichens and microorganisms, are known for their ability to specifically bind to carbohydrates. Plant lectins can be classified into 12 families including the Galanthus nivalis agglutinin (GNA)-related lectin superfamily, which is widespread among monocotyledonous plants and binds specifically to mannose, a behavior that confers remarkable anti-tumor, anti-viral and insecticidal properties on these proteins. The present study characterized a mitogenic lectin from this family, called tarin, which was purified from the crude extract from taro (Colocasia esculenta). The results showed that tarin is a glycoprotein with 2-3% carbohydrate content, composed of least 10 isoforms with pIs ranging from 5.5 to 9.5. The intact protein is a heterotetramer of 47kDa composed of two non-identical and non-covalently associated polypeptides, with small subunits of 11.9kDa and large subunits of 12.6kDa. The tarin structure is stable and recovers or maintains its functional structure following treatments at different temperatures and pH. Tarin showed a complex carbohydrate specificity, binding with high affinity to high-mannose and complex N-glycans. Many of these ligands can be found in viruses, tumor cells and insects, as well as in hematopoietic progenitor cells. Chemical modifications confirmed that both conserved and non-conserved amino acids participate in this interaction. This study determined the structural and ligand binding characteristics of a GNA-related lectin that can be exploited for several different purposes, particularly as a proliferative therapeutic molecule that is able to enhance the immunological response.

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“…In a search for this fraction, LLGP was subjected to reduction and alkylation treatments prior to biochemical and immunological assays. Thereafter GP39-42 and GP24 were also exhaustively reduced and alkylated in 8 M urea/0.6 M TRIS-HCl (pH 8.6) for 2 h. Each sample was mixed with 4 M dithiothreitol and incubated in N 2 for 30 min and then in 500 mM iodoacetamide solution for 30 min at 37°C (Aitken and Learmonth 1996).…”

Section: Antigen Puri®cationmentioning

confidence: 99%

Characterization of GP39-42 and GP24 antigens from Taenia solium cysticerci and of their antigenic GP10 subunit

Plancarte

Hirota

Martinez-Ocaña

et al. 1999

Parasitology Research

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Diagnosis of neurocysticercosis is performed by Western blotting with an enriched fraction of glycoproteins (LLGP). GP39-42 and GP24 are immunodominant antigens. These antigens were electroeluted and characterized by biochemical methods. When GP39-42 or GP24 were reduced, a band of 10 kDa (named GP10) appeared; this band was also analyzed. The most abundant amino acids in the three GPs were lysine, phenylalanine, asparagine, glycine, and leucine. The amino terminal portion was sequenced, and the following order was obtained for the three GPs: EKNKPKNVAXSTKKGYEYVXEF. The glycan portion was 8.4%, 18.2%, and 18.3% in GP39-42, GP24, and GP10, respectively. The three GPs contained mannose, N-acetyl-D-glucosamine, and galactose. GP39-42 and GP24 seem to be oligomeric forms of GP10. When reduced LLGP was reacted with samples obtained from patients with neurocysticercosis or pigs with cysticercosis, a band corresponding to GP10 was always observed. Furthermore, hyperimmune serum from rabbits immunized with GP39-42 or with GP24 recognized GP10 as well as GP39-42 and GP24. The data obtained in this paper suggest that GP10 might be a useful tool for diagnosis.

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Carboxymethylation of Cysteine Using Iodoacetamide/ Iodoacetic Acid

Cited by 20 publications

References 0 publications

Properties and Mutation Analysis of the CelK Cellulose-Binding Domain from the Clostridium thermocellum Cellulosome

Properties and Mutation Analysis of the CelK Cellulose-Binding Domain from the Clostridium thermocellum Cellulosome

Structural analysis and binding properties of isoforms of tarin, the GNA-related lectin from Colocasia esculenta

Characterization of GP39-42 and GP24 antigens from Taenia solium cysticerci and of their antigenic GP10 subunit

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