Primary Structure of Bovine Growth Hormone

Santomé, J.A.; Dellacha, Juan M.; Paladini, Alejandro C.; Peña, Clara; Biscoglio, Mirtha J.; Daurat, S.T.; Poskus, Edgardo; Wolfenstein, C.E.M.

doi:10.1111/j.1432-1033.1973.tb02971.x

Cited by 53 publications

(16 citation statements)

References 36 publications

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“…b-STH is a single-chain protein hormone consisting of 191 amino acids. The native structure of rb-STH consists of a four-helix bundle motif with two intrachain disulfide bridges at positions 153/164 and 181/189 (13,14). Physical instability due to perturbation ofthe tertiary structure results in partially unfolded intermediates and potential formation of insoluble aggregates (15).…”

Section: Introductionmentioning

confidence: 99%

Controlled release of polypeptides from polyanhydrides.

Ron

Turek

Mathiowitz

et al. 1993

Proc. Natl. Acad. Sci. U.S.A.

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The effects of both polymer hydrophobicity and addition of stabilizers on the release and integrity of polymer-encapsulated proteins were studied. By using very hydrophobic poly [l,3-bis(p-carboxyhydroxy)hexane anhydride] with sucrose as an excipient, both recombinant bovine somatotropin and zinc insulin were released intact over 3 weeks. The released proteins appeared to maintain their integrity as judged by acidic reverse-phase HPLC, size-exclusion HPLC, radioimmunoassay, and conformation-sensitive immunoassays. Our results also suggest how polymer hydrophobicity can be used to enhance protein stability. more important is the chemical instability of rb-STH in aqueous environments leading to deamidated, chain-cleaved, and covalently bonded oligomeric products (16). This instability presents an intriguing problem to attempt to overcome; solving it will demonstrate the potential of using hydrophobic polymers to reduce aggregation and to control the release of unstable proteins. Insulin was used to study the extent to which in vitro release and activity would compare with in vivo activity. The diabetic rat model is well documented in the literature and the successful use of our final formulation for delivery of intact protein would be a good affirmation.Most peptidic and proteinic drugs are susceptible to degradation at the site of administration, whether administered by subcutaneous, intramuscular, intestinal, buccal, rectal, nasal, or vaginal routes (1, 2). In addition, some proteins have very short in vivo half-lives and, as a consequence, multiple injections are required to achieve desirable therapy. One way to increase the therapeutic efficacy of these proteins is to incorporate them in a controlled release system that releases the proteins continuously. Several controlled release systems have been tested for their ability to sustain the release of peptidic drugs such as insulin (3), epidermal growth factor (4), interferon (5), and luteinizing hormone-releasing hormone and its analogs (6-9). It is also possible that, in addition to providing sustained release, polymers may help to protect the polypeptides from denaturing conditions, water, and proteolytic enzymes. Furthermore, the use of bioerodible polymers does not require posttreatment removal of the device. However, the polymers used for such delivery are limited to those that are inert toward the incorporated proteins-i.e., those polymers that neither bind to irreversibly nor promote decomposition of the proteinaceous therapeutic agent. Polyanhydrides have been investigated in our and other laboratories as biodegradable implants (10-12). Furthermore, polyanhydrides are one of the few synthetic biodegradable polymers being studied clinically as injectable or as implantable matrices. We used several polyanhydrides with different hydrophobicities to explore the effect of hydrophobicity and excipients on the release and activity of two proteins. We also studied the implications of possible interactions between the polymer, its degradation products, a...

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Section: Introductionmentioning

confidence: 99%

Controlled release of polypeptides from polyanhydrides.

Ron

Turek

Mathiowitz

et al. 1993

Proc. Natl. Acad. Sci. U.S.A.

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“…Second, there is a relatively hydrophobic stretch of 24 amino acids at the start of this variant region, delineated by an arginine at the end of exon 4 and a glutamic acid at amino acid residue 148, raising the possibility that this polypeptide might be located in a membrane. Third, in wild-type growth hormone, there exists a disulfide bond between cysteine-53 and cysteine-164 (22). Cysteine-164 would be eliminated in a polypeptide derived from the intron D-containing mRNA.…”

Section: Discussionmentioning

confidence: 99%

Alternative processing of bovine growth hormone mRNA: nonsplicing of the final intron predicts a high molecular weight variant of bovine growth hormone.

Hampson¹,

Rottman²

1987

Proc. Natl. Acad. Sci. U.S.A.

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We have detected a variant species of bovine growth hormone mRNA in bovine pituitary tissue and in a stably transfected bovine growth hormone-producing cell line. Analysis of this variant mRNA indicated that the last intervening sequence (intron D) had not been removed by splicing. Inspection of the sequence of intron D reveals an open reading frame through the entire intron, with a termination codon encountered 50 nucleotides into the fifth exon, which is shifted from the normal reading frame in this variant mRNA. If translated, this variant mRNA would encode a growth hormone-related polypeptide having 125 amino-terminal amino acids identical to wild-type growth hormone, followed by 108 carboxyl-terminal amino acids encoded by the 274 bases of intron D along with the first 50 nucleotides of exon 5. This variant polypeptide would be 42 amino acids longer than wild-type bovine growth hormone or approximately 5000 greater in molecular weight. The intron D-containing variant of bovine growth hormone mRNA was demonstrated to exist on polysomes, suggesting that this mRNA species is translated into a polypeptide. Cytosolic mRNA species containing any of the other three introns of the bovine growth hormone gene were not detectable.

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“…Amino acid sequences for bSt which differed slightly were published almost simultaneously by Santome et al (1973) and Wallis (1973) . Miller et al (1980) confirmed the Wallis sequence ( Fig.…”

Section: Bst Primary Structurementioning

confidence: 99%

“…Pituitaryderived bSt has been reported to show sequence heterogeneity at the NH zterminus where approximately one-half of the pituitary bSt molecules contain a full-length sequence beginning with Ala-Phe-Pro-Ala-Met-. The other half ofthe molecules begins with Phe-Pro-Ala-Met- (Li and Ash, 1953;Santome et al, 1973;Wallis, 1973). This heterogeneity is likely due to variability in the removal of a prehormone leader sequence (Lingappa et al, 1977).…”

Section: Bst Primary Structurementioning

confidence: 99%