. Asiatic herpetological research. Reptiles -- Asia Periodicals; Amphibians -- Asia Periodicals. April 1990 Asiatic Herpetological Research Vol. 3, p. 117 100 o CM C\J <. 40 60 time(min) FIG. 1. RP-HPLC of the water eluate from the alumina columns. Column: uBondapak C\% mm. Mobile phase: A= CF3COOH B=60% CH3CN in A. Concave gradient elution from 0-85%B (dotted line), 90 min, at ml/min. Detected at UV 220 nm, aufs. Gly-Leu-Val-Ser-Asp-Leu-Met-Tyr-Gly-Ile-Gly-Leu-NH2 â GH-1- A h â CH-2- FlG. 2. Profile of sequence anlaysis of the dodecapeptide. (-*): DABITC/PITC method. (
. Asiatic herpetological research. Reptiles -- Asia Periodicals; Amphibians -- Asia Periodicals. April 1990 Asiatic Herpetological Research Vol. 3, p. 117 100 o CM C\J <. 40 60 time(min) FIG. 1. RP-HPLC of the water eluate from the alumina columns. Column: uBondapak C\% mm. Mobile phase: A= CF3COOH B=60% CH3CN in A. Concave gradient elution from 0-85%B (dotted line), 90 min, at ml/min. Detected at UV 220 nm, aufs. Gly-Leu-Val-Ser-Asp-Leu-Met-Tyr-Gly-Ile-Gly-Leu-NH2 â GH-1- A h â CH-2- FlG. 2. Profile of sequence anlaysis of the dodecapeptide. (-*): DABITC/PITC method. (-»): gas phase sequencing. («-): CP-Y digestion. CH: a-chymotryptic peptides. difference of retention time of the two peaks was min. The peak 32 acted as the representative of the dodecapeptide for further studies and the peak 33 was also investigated simultaneously. Amino acid composition of the peak 32 was Asp (1), Ser (l),Gly (3), Val (1), Met (1), He (1), Leu (3), Tyr (1). Amino acid sequence of the peak 32 was determined by the DABITC/PITC method and gas phase sequencing. The former method proceeded to the tenth step, but the latter to the penultimate residue (Fig. 2). For C- terminal residue analysis, CP-A and CP-Y digestions of the peak 32 were carried out, and did not release any amino acids by the former. This indicated a blocked C- terminus; upon latter, however, Leu and Gly were obtained. Thus, we deduced that the C-terminal structure of the peak 32 is Leu-NH2. Digestion of the peptide by a- chymotrypsin provided further structural confirmation. The fragment peptides, CH- 1 and CH-2 were separated on HPLC as depicted in Fig. 3. Amino acid compositions of the two fragments are in accordance with their sequences (see Fig. 2), respectively. From the above results the complete amino acid sequence of the peak 32 was. Please note that these images are extracted from scanned page images that may have been digitally enhanced for readability - coloration and appea
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