W. G. Kruggel

A complete amino acid sequence for the basic subunit of crotoxin

The complete amino acid sequence of the basic subunit of crotoxin from the venom of Crotalus durissus terrificus has been determined. Fragmentation of the protein was achieved by using cyanogen bromide and arginine- and lysine-specific endoproteases. Sixteen Glx and Asx residues reported by Fraenkel-Conrat et al. (1980) in Natural Toxins (D. Eaker and T. Wadstrom, eds.), pp. 561-567, Pergamon, Oxford.) have been resolved as Glu or Gln and Asp or Asn residues, respectively. Most of the remaining sequence is identical to that reported by the foregoing authors although several significant differences were evident in our protein. Tyr-61 was not present; thus the correct sequence is Lys-60, Trp-61. The latter sequence aligns with sequences of all other known viperid and crotalid phospholipases A2 (S. D. Aird, I. I. Kaiser, R. V. Lewis, and W. G. Kruggel (1985) Biochemistry 24, 7054-7058). Other differences include Asx-99, which is Ser, and Asx-105, which is Tyr. Some positions display allelic variation. In some lots of venom Glx-33 is Gln, while in others it is Arg. Positions 37 and 69 occur as mixtures of both Lys and Arg. Amino acid sequence comparisons between the basic and acidic subunits of crotoxin and between the basic subunit and other phospholipase A2 molecules indicate that the basic subunit is structurally most similar to the monomers of nontoxic, dimeric phospholipases A2 from the venoms of Crotalus adamanteus, Crotalus atrox, and Trimeresurus okinavensis, and to the toxic monomeric phospholipase A2 from the venom of Bitis caudalis.

The amino terminal sequences of bovine and human chromogranin A and secretory protein I are identical.

The amino terminal sequences of bovine and human adrenal medullary chromogranin A have been determined. Their sequences are identical and also identical to the published sequence of secretory protein I from the parathyroid gland. This data indicates that the previously published sequence of chromogranin A is incorrect at residues 2 and 19. These data confirm earlier observations of a substantial similarity between secretory protein I and chromogranin A and, in fact, strongly suggest that they are identical.

Amino acid sequence of the basic subunit of mojave toxin from the venom of the mojave rattlesnake (Crotalus S. scutulatus)

The complete sequence of the basis subunit of Mojave toxin from the venom of the Mojave rattlesnake (Crotalus s. scutulatus) is presented. It is shown to have great similarity to the basic subunits of related toxins from the venoms of the South American and midget faded rattlesnakes.

Multiple myotoxin sequences from the venom of a single prairie rattlesnake (Crotalus viridis viridis)

Multiple myotoxin a sequences have been determined from the venom of a single adult male prairie rattlesnake (Crotalus viridis viridis). This is the first time such individual variation has been reported for this toxin class and the number of isoforms suggest that myotoxin a is the product of a duplicated locus.

Purification and sequence of a novel ovine adrenal medullary peptide and its precursor.

A 24 amino acid polypeptide that does not originate from (pre)proenkephalin has been isolated from ovine adrenal chromaffin granules. Its sequence is: Arg-Leu-Pro-Gly-Glu-Leu-Arg-Asn-Tyr-Leu-Asp-Tyr-Gly-Glu-Glu-Val-Gly-Glu- Glu-Ala -Ala-Arg-Gly-Val. This peptide is generated from a precursor molecule that has also been purified and partially sequenced. The proteolytic cleavage occurs at a triple Arg site. A search of the available protein sequence data banks shows very little homology to any known protein.

Purification and sequence of an opioid peptide derived from ovine proenkephalin.

An enkephalin-containing peptide originating from ovine adrenal proenkephalin has been purified and sequenced. The sequence of the peptide is: GLY-GLY-GLU-VAL-LEU-GLY-LYS-ARG-TYR-GLY-GLY-PHE-MET (preproenkephalin 128-140) which represents a portion of peptide F (preproenkephalin 107-140). This peptide has a sequence identical to that of bovine preproenkephalin 128-140 while it differs from the corresponding human sequence in positions 129, 131 and 133.

Purification and sequence of a non-opioid peptide derived from ovine proenkephalin: implications for possible species specific processing.

A non-enkephalin containing pentadeca peptide derived from ovine adrenal proenkephalin has been purified and sequenced. The sequence of the peptide is: Phe-Ala-Glu-Pro-Leu-Pro-Ser-Glu-Glu-Glu-Gly-Glu-Ser-Tyr-Ser (preproenkephalin 237-251) representing the amino portion of peptide B (preproenkephalin 237-268). The sequence is identical to bovine preproenkephalin 237-251, differing from the corresponding human sequence at positions 240 and 244. This peptide can be generated by a processing event common to other opioid peptides and is present in chromaffin granules in significant amounts. The presence of this peptide in substantial quantities suggests a possible difference in proenkephalin processing between the bovine and ovine adrenal medulla.

Presence of pancreatic trypsin inhibitor in adrenal medullary chromaffin cells

A bovine pancreatic trypsin inhibitor was isolated from bovine adrenal medullary chromaffin granules. Its N-terminal sequence is: arg-pro-asp-phe-cys-leu-glu-pro-pro-tyr-thr-gly-pro-cys-lys-ala-arg-ile- arg-tyr- phe-tyr-asn-ala-lys-ala-gly-leu-cys-gln-thr-phe-val-tyr-gly-gly-cys-arg- ala-lys-arg-asn-asn-phe-lys- which corresponds precisely with the N-terminus of Bovine Pancreatic Trypsin inhibitor. The presence of this inhibitor in these granules suggests another method of regulating the prohormone proteases present there.