ABSTRACT
5-Bromo-2[(2-iodoacetyl)amino]benzenesulfonic acid (AIBSA), a reagent for modification of free of cysteine thiol groups in proteins and peptides, was synthesized. Rate constants of its interaction with thiol groups were determined. The presence of a bromine atom allows an easy identification of the AIBSA-labeled peptides in mass spectra due to the characteristic isotope distribution. The compound is stable in solution and under exposure to light.
Subject(s)
Benzenesulfonates/chemistry , Bromine/chemistry , Cysteine/chemistry , Iodoacetamide/analogs & derivatives , Proteins/chemistry , Benzenesulfonates/chemical synthesis , Benzenesulfonates/radiation effects , Iodoacetamide/chemical synthesis , Iodoacetamide/chemistry , Iodoacetamide/radiation effectsABSTRACT
Sequencing of fragments of genes coding for silicic acid transport (SIT) proteins of diatoms of evolutionary distant classes (centric Chaetoceros muelleri Lemmermann, pennate araphid Synedra acus Kützing, pennate raphid Phaeodactylum tricornutum Bohlin, and pennate with keeled raphe system Cylindrotheca fusiformis Reimann et Lewin), revealed the presence in these proteins of a conservative amino acid motif CMLD. Hydropathy profiles suggest that CMLD occupies a position between two transmembrane strands which do not contain lysine and arginine residues. The two strands are good candidates for the role of the channel along which transport of silicic acid occurs. CMLD is a rare motif. Diatoms are known to need Zn2+ for the incorporation of silica. Presumably, CMLD is the site of Zn2+ binding of SITs. We found that the growth of diatoms is inhibited by a negatively charged alkylating reagent 5-(2-iodoacetamidoethyl)aminonaphtalene-1-sulfonic acid which cannot penetrate through the cell membrane. Cysteine of CMLD can be a target of this reagent. Synthetic peptide NCMLDY forms a complex with Zn2+, as revealed by the fact that the ion considerably reduces the rate of alkylation of the peptide.
