Purification of somatostatin from frog brain: coisolation with retinal somatostatin-like immunoreactivity.
Somatostatin-like immunoreactivity (SLI) was purified from frog brain and retina, and the structure of the brain peptide was determined. Frog brain (101 g) and retinal (45 g) tissues were extracted with 3% acetic acid, yielding 9.6 and 0.44 nmol of SLI, respectively. SLI was further purified by chromatography on a somatostatin immunoaffinity column followed by sequential application to reverse-phase C-18 HPLC columns. The brain and retinal peptides, purified roughly 100,000-fold with net yields of 7.5 and 2.3%, respectively, appeared identical in the final steps of purification. The amino acid sequence of brain SLI, as determined by a gas-phase automated Edman degradation technique, was as follows: Ala-Gly-(Cys)-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-(Cys). Our data indicate that despite structural variations in somatostatins of other lower vertebrates, the amino acid sequence of frog brain and, by deduction, retinal SLI is identical to that of somatostatin tetradecapeptide. These findings support the physiological relevance of studies directed at elucidating the neurotransmitter function of somatostatin using the well-established models of frog brain and retina.
Takami
M
M
Reeve
J R
JR
Hawke
D
D
Shively
J E
JE
Basinger
S
S
Yamada
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000502:physiology*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D005399:Fishes; D011863:Radioimmunoassay; D011894:Rana pipiens; D012160:Retina / Q000502:physiology*; D013004:Somatostatin / Q000235:genetics / Q000302:isolation & purification*
D013004:Somatostatin