Peptides of a biologically active tryptic digest of bovine growth hormone

Yamasaki, Nobuyuki; Kikutani, Motosuke; Sonenberg, Martin

doi:10.1021/bi00807a009

Cited by 62 publications

(28 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The smaller peptide, given in large doses to two pituitary dwarfs, induced some nitrogen retention and had a marked anti-insulin effect. This peptide is homologous in sequence with peptide 95-134 of human growth hormone (12).…”

Section: Introductionmentioning

confidence: 98%

“…Sonenberg et al (5) in 1965 described partial digestion of bGH with trypsin; they used soy bean trypsin inhibitor to control the extent of digestion. The purification and characterization of the reaction products have been described in a series of papers (6)(7)(8)(9)(10)(11)(12). Briefly digested bGH, in which 2-3 peptide bonds were cleaved, is completely active in the rat (tibia test), and in hypopituitary human subjects at doses of 12-100 mg/day causes nitrogen retention, free fatty acid mobilization and impaired glucose tolerance.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Metabolic Effects of Plasmin Digests of Human Growth Hormone in the Rat and Man

Mills¹,

Reagan²,

Rudman³

et al. 1973

J. Clin. Invest.

View full text Add to dashboard Cite

A B S T R A C T As a first step in our study of structurefunction relationships among primate and non-primate growth hormones, human growth hormone (hGH) was subjected to the limited digestive activity of human plasmin. The lyophilized whole digest, containing less than 2% of unchanged hormone, had an average of 2. The digest also caused glucosuria in partially pancreatectomized rats treated with dexamethasone.These metabolic actions of plasmin-digested hGH in the array of animal tests were confirmed by comparable effects elicited in 11 human subjects (nine pituitary-deficient children and adolescents and two nondeficient adults). A single injection of the plasmin digest caused an increase in plasma free fatty acids and a fall in plasma amino acids. Seven daily injections caused positive balances of nitrogen, phosphorous, sodium, and potassium, gain in body weight, and in two of three subjects impairment of glucose tolerance. The potency of the plasmin digest in producing these metabolic effects in man was comparable to that of native hGH.Thus, 2-3 bonds in the hGH molecule can be cleaved by plasmin without impairing the hormone's growth-

show abstract

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Metabolic Effects of Plasmin Digests of Human Growth Hormone in the Rat and Man

Mills¹,

Reagan²,

Rudman³

et al. 1973

J. Clin. Invest.

View full text Add to dashboard Cite

show abstract

“…The biologically active fragment (A-II) obtained from a tryptic digest of BGH was prepared by the method of Yamasaki et al (14). [This A-II fragment of molecular weight 5000 and 37 amino acids was homogeneous by polyacrylamide disc electrophoresis, gel filtration, and sedimentation equilibrium (14).] All other reagents were of analytic grade.…”

Section: Methodsmentioning

confidence: 99%

“…BGH was prepared in this laboratory by a modification (12) of the method of Dellacha and Sonenberg (13), and was homogeneous by polyacrylamide disc electrophoresis. The biologically active fragment (A-II) obtained from a tryptic digest of BGH was prepared by the method of Yamasaki et al (14). [This A-II fragment of molecular weight 5000 and 37 amino acids was homogeneous by polyacrylamide disc electrophoresis, gel filtration, and sedimentation equilibrium (14).]…”

mentioning

confidence: 99%

Interaction of Human Growth Hormone and Human Erythrocyte Membranes Studied by Intrinsic Fluorescence

Sonenberg

1971

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

Self Cite

View full text Add to dashboard Cite

The intrinsic fluorescence of hunman erythrocyte membranes excited by unpolarized and polarized light has been studied with and without the addition of human growth hormone. The peak emission intensity of the membranes appeared at 332 nm, with a distinct shoulder at about 303 nm. Excitation spectra contained two peaks, at 282 and 225 nm. Fluorescence polarization was maximal (about 0.4) The data are consistent with the proposition that human growth hormone, by some cooperative mechanism, produces a conformational change in the membrane proteins with associated depolarization of fluorescence.A change was noted in the far-ultraviolet circular dichroism (CD) spectrum of human erythrocyte membranes in the presence of human growth hormone (HGH), but not of bovine hormone (BGH) (1). In addition, the near-ultraviolet CD spectrum of HGH was altered by human erythrocyte membranes (1), suggesting a change in the conformation of the hormone. It was considered likely that the changes in optical activity of the membranes were not the result of distortions or artifacts of particulate (or membrane) systems (2-6), since no changes in light scattering or phase-contrastmicroscopic appearance of the membranes were noted (1). This suggested that the decreased optical activity was the consequence of conformational changes in membrane proteins upon interaction with HGH. In order to obtain more data about possible conformational changes of membrane proteins, the interaction of erythrocyte membranes and HGH has been studied by fluorescence and polarization of intrinsic fluorescence. MATERIALS AND METHODSSterile solutions were used to prepare erythrocyte membranes from freshly drawn human blood by methods detailed by Dodge et al. (7), Marchesi et al. (8), or Steck et al. (9). Stock suspensions of these membrane preparations were maintained at 4°C and were used as long as they were responsive to HGH. Protein concentrations of the membrane preparations were determined from the A2m of 2-chloroethanol solutions prepared from measured amounts of the original membrane suspension (10). Membranes were counted under a phasecontrast microscope in a Petroff-Hauser counter. HGH was prepared from human pituitaries obtained at autopsies by the method of Saxena and Henneman (11), and contained one major and two minor bands by polyacrylamide disc electrophoresis. BGH was prepared in this laboratory by a modification (12) of the method of Dellacha and Sonenberg (13), and was homogeneous by polyacrylamide disc electrophoresis. The biologically active fragment (A-II) obtained from a tryptic digest of BGH was prepared by the method of Yamasaki et al. (14). [This A-II fragment of molecular weight 5000 and 37 amino acids was homogeneous by polyacrylamide disc electrophoresis, gel filtration, and sedimentation equilibrium (14).] All other reagents were of analytic grade.Fluorescence measurements were made on a Cary 50-026-900 differential spectrophotofluorometer employing frontsurface illumination, where the exciting and emitting beams subtend an an...

show abstract

“…If we assume that four methionine residues involved in bGH1.7) were completely labeled with iodoacetamide spin label [IJ, the composite ESR specttrum can be explained by a hypothesis that these residues are not necessarily involved in a uniform environment but some of them are in the region where the spin label is strongly immobilized and others in the region where the spin label is only slightly, if at all, immobilized. Recently, Chen and Sonenbeberg 10 ) predicted a conformation of bGH by the ChouFassman methodY-16) According to their prediction, methionine at 5 exists in the random coil region occurring at the amino terminal portion of bGH (1)(2)(3)(4)(5)(6)(7)(8)(9) and methionine at 124 in the a-helical region (111-127). Two methionine residues at 149 and 179 are involved in the regions of fl-turn (146-149) and Jl-sheet structure (174-179), respectively.…”

Section: Received June 13 1979mentioning

confidence: 99%