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[The role of single amino acid residues of the immunodominant continuous epitope of cytochrome P450CAM (CYP101) 312LKKGDQ317 in recognition by specific antibodies].

As it has been shown previously the site 311-318 of bacterial cytochrome P450cam (CYP101) contains an immunodominant continuous B-epitope. In order to investigate the role of single amino acid residues in antibody binding antigenic hexapeptide 312LKKGDQ317 analogues including single amino acid replacements were synthesized using Multipin technology. Antibodies from three anti-P450cam polyclonal rabbit sera interacted similarly to these peptides. The residue G315 was found to play a significant role in antibody recognition; any replacement leads there to considerable decrease of antibody binding. Residues L312, K313 and D316 occurred to be partly replaceable, whereas K314 and Q317 were not essential in recognition. These results correspond to known spatial structure of P450cam molecule. In its 312-317 site the polypeptide chain makes a turn and so some water-accessible atoms form a compact surface cluster including side chain atoms of K313 and D316, O-atom of K314 and C alpha-atom of G315, which present reliable Ig binding site. Side chain of K314 outlying from this cluster does not participate in the interaction. The received data permit to consider that the continuous epitope 312-317 of P450cam is conformationally dependent.

Amino Acids↗
PubMed1999

[The enantiomeric separation of aromatic alcohol amino drugs by thin-layer chromatography].

Two chiral aromatic alcohol amino drugs, Labarol and Bataroc, were resolved by thinlayer chromatography (TLC) on the silica gel GF254 plates (2.5 cm x 10 cm), by using the ammonium-D-10-camphorsulfonate (CSA) as chiral ion-pair interaction agent which was added to the mobile phase in the ammonium form. All developments were carried out at lower temperature (2-4 degrees C, in a refrigerator) in small glass jars of 250 mL volume. These two drugs were not resolved at room temperature (15-30 degrees C). Analytical reagent grade methanol and dichloromethane can be directly used as mobile phase without further drying. The chiral separation occured over a range from 40% to 70% (optimum 67%) dichloromethane in the mobile phase volume ratio and 55% to 80% (optimum 60%) dichloromethane in the mobile phase volume ratio. These separation conditions were easily obtained. This method is relatively inexpensive and attractive.

Amino Alcohols↗