Carp Insulin: Amino Acid Sequence, Biological Activity and Structural Properties

Makower, Alexander; Dettmer, R; Rapoport, Tom A.; Knospe, S; Behlke, Joachim; Prehn, Siegfried; Franke, P.; Etzold, G.; Rosenthal, S

doi:10.1111/j.1432-1033.1982.tb05886.x

Cited by 24 publications

(8 citation statements)

References 26 publications

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“…The data indicate that carp preproinsulin contains 108 amino acid residues, 21 in the signal peptide, 31 in the B-chain, 35 in the C-peptide region and 21 in the A-chain. The deduced primary sequence of carp insulin is in agreement with the protein sequence determined by Makower at al (2). The molecular weight of carp preproinsulin of 11 800 D, calculated on the basis of the sequencing data, is identical with that deduced by immunoprecipitation of preproinsulin after cellfroo translation (19,20 to be due to a shift of the cleavage site of the signal peptidase by two amino acid residues towards the N-terminus (see fig.…”

Section: Introductionsupporting

confidence: 71%

“…4541 acid sequence derived from it. To verify the nucloic acid sequencing data the carp insulin amino acid sequence has been estimated independently and already reported (2). The knowledge of the preproinsulin sequence of carp (Cyprinus carpio), a fresh water fish, allows a valuable extension of the comparisons of evolutionary distant vertebrate insulin genes.…”

Section: Introductionmentioning

confidence: 99%

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Carp preproinsulin cDNA sequence and evolution of insulin genes

Hahn¹,

Winkler²,

Rapoport³

et al. 1983

Nucl Acids Res

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Section: Introductionsupporting

confidence: 71%

Section: Introductionmentioning

confidence: 99%

Carp preproinsulin cDNA sequence and evolution of insulin genes

Hahn¹,

Winkler²,

Rapoport³

et al. 1983

Nucl Acids Res

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“…It has been suggested by Makower et al [39] that a charge compensation between residues B,, and B,, might be a necessary feature to attain a reasonable biological response. However, such a charge compensation does not exist for dogfish insulin (B,,Pro, B,,Lys) and yet we observe a potency that is exhibited by teleost insulins in general.…”

Section: Discussionmentioning

confidence: 99%

“…Pro has been substituted for Glu at B,, as seen in carp insulin [39] and Lys substituted for Arg at Bz2.…”

Section: Discussionmentioning

confidence: 99%

Dogfish insulin

Bajaj

Blundell

Pitts

et al. 1983

European Journal of Biochemistry

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Insulin from an elasmobranch, the spiny dogfish (Squalus acantlzias) has been purified to near homogeneity by means of acid-ethanol extraction and salt precipitation. The amino acid sequences of the performic-acid-oxidised A and B chains have been determined and exhibit some unusual features. The A chain contains a total of 22 amino acids; only the insulin from coypu (a member of the Rodentia suborder, Hystricomorpha), has previously been reported to contain an extension past the A,, asparagine. The B,, histidine, which is involved in the formation of the insulin hexamers in higher vertebrates through the co-ordination of zinc, is present in this elasmobranch insulin. Several substitutions relative to bovine insulin occur in the proposed receptor binding region (A,Gln+ His, B, Glu +Pro, B,,Arg+Lys, B,,Phe+Tyr). In spite of these substitutions, the maximal response in the rat epididymal fat cell assay is the same for bovine and dogfish insulins; the concentration required to produce the half-maximal response is, however, approximately threefold greater for dogfish insulin than that of bovine insulin.The use of interactive computer graphics model-building predicts that the dogfish insulin can attain a threedimensional structure very similar to that of bovine insulin; circular dichroic spectra are presented which support the model-building studies.In the years since the determination of the amino acid sequence of bovine insulin [I] and the subsequent elucidation of the three-dimensional structure of the protein hormone [2], many insulins from different species have been studied. These investigations have attempted to corrclate the physical and biological properties of the hormone to specific amino acid residues in the sequence [3,4], as well as to understand thc evolution of the hormone [5,6].Immunocytochemical and biological studies suggest the presence of an insulin-like activity in prokaryotes such as Escherichia coli K12 [7], in various unicellular eukaryotes [S], in the protostomian [9] and the deuterostomian [lo] lines of invertebrate evolution [5]. However, the complete chemical characterization of these insulin immunoreactivities has not yet been possible. Though representative insulin sequences from teleosts reptiles, aves and mammals, are available, 21 lack of information exists at the level of lowermost gnathastomian vertebrates, namely the cartilaginous fish [5].Insulin from the hagfish (Myuine glutinosa) a jawless fish, is the phylogenically most original insulin to be scqucnced [5,1 I]. X-ray analysis [I21 suggests that this insulin has a spatial structure very similar to porcine insulin, even though the cyclostomian line seems to have diverged from the main line of vertebrate evolution nearly 500 million years ago [13]. Hagfish insulin has a very low biological potency when assayed in mammalian systems. Further it lacks the histidine at B,, which is responsible for the formation of zinc-insulin hexamers in most mammalian insulins. In contrast, several teleost insulins that have been studied can for...

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“…These changes are responsible for their generally poor cross-reactivity with antibodies to mammalian insulin [4], but have a much less dramatic effect on their biological potency which is about 30 -50% that of mammalian insulins in mammalian test systems [l, 51. The ability to form dimers and probably also zinc-containing hexamers is apparent from the amino acid sequences and some supporting evidence is available for the latter [5,6]. However, as yet, there is no detailed three-dimensional structural information available on a teleost fish insulin, aside from model building predictions, although the crystal structure of the more primitive hagfish, a cyclostome, has been determined [7].…”

mentioning

confidence: 99%

The isolation, purification and amino‐acid sequence of insulin from the teleost fish Cottus scorpius (daddy sculpin)

Cutfield

Carne

et al. 1986

European Journal of Biochemistry

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Insulin from the principal islets of the teleost fish, Cottus scorpius (daddy sculpin), has been isolated and sequenced. Purification involved acid/alcohol extraction, gel filtration, and reverse-phase high-performance liquid chromatography to yield nearly 1 mg pure insulin/g wet weight islet tissue. Biological potency was estimated as 40% compared to porcine insulin. The sculpin insulin crystallised in the absence of zinc ions although zinc is known to be present in the islets in significant amounts. Two other hormones, glucagon and pancreatic polypeptide, were copurified with the insulin, and an N-terminal sequence for pancreatic polypeptide was determined.The primary structure of sculpin insulin shows a number of sequence changes unique so far amongst teleost fish. These changes occur at A14 (Arg), A15 (Val), and B2 (Asp). The B chain contains 29 amino acids and there is no N-terminal extension as seen with several other fish. Presumably as a result of the amino acid substitutions, sculpin insulin does not readily form crystals containing zinc-insulin hexamers, despite the presence of the coordinating B10 His.Sequence studies on a wide range of vertebrate insulins have been helpful in conjunction with the known three-dimensional structure of pig insulin in explaining physical, chemical and biological properties of the hormone [l, 21. Insulin molecules from the teleostii (bony fish) display characteristic sequence changes in comparison to mammalian insulins, especially in the central region of the A chain and also in the N-terminal region of the B chain [3]. These changes are responsible for their generally poor cross-reactivity with antibodies to mammalian insulin [4], but have a much less dramatic effect on their biological potency which is about 30 -50% that of mammalian insulins in mammalian test systems [l, 51. The ability to form dimers and probably also zinc-containing hexamers is apparent from the amino acid sequences and some supporting evidence is available for the latter [5,6]. However, as yet, there is no detailed three-dimensional structural information available on a teleost fish insulin, aside from model building predictions, although the crystal structure of the more primitive hagfish, a cyclostome, has been determined [7].

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Carp Insulin: Amino Acid Sequence, Biological Activity and Structural Properties

Cited by 24 publications

References 26 publications

Carp preproinsulin cDNA sequence and evolution of insulin genes

Carp preproinsulin cDNA sequence and evolution of insulin genes

Dogfish insulin

The isolation, purification and amino‐acid sequence of insulin from the teleost fish Cottus scorpius (daddy sculpin)

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