The amino acid sequence of bovine growth hormone.
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ABSTRACT: Reactivity of histidines in bovine growth hormone towards ethoxyformic anhydride was investigated and localization in the molecule of two kinetically distinguishable classes was achieved, a slow class including only histidine residue 169 (k = 0.180 min-1) and a fast one composed of histidines 19 and 21 (k = 0.900 min-1). Total ethoxyformylation of bovine growth hormone brought about a complete loss of its capacity to compete with 125I-labelled hormone for rat-liver binding sites, but modification of approximately half of the fast histidine group was enough to produce an important decrease in this capacity. Circular dichroism studies indicated no significant changes in protein conformation with all three histidine residues modified. Practically full binding capacity was restored when these residues were regenerated by treatment with hydroxylamine. These results suggest that one or both of the fast reacting histidine residues are involved in bovine growth hormone binding to its specific receptors.International journal of peptide and protein research 05/1983; 21(4):451-7.
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ABSTRACT: A nonapeptide, Arg-Glu-Leu-Glu-Asp-Gly-Thr-Pro-Arg, corresponding to the 123–131 sequence of bovine pituitary growth hormone, was prepared by a modified solid phase synthesis. To avoid an acid- or base-catalysed rearrangement at the Asp-Gly bond, which gives rise to an aspartimide-containing peptide, a new combination of protecting groups was introduced. The 2-(4-biphenylyl)-2-propyloxycarbonyl group was used for Nα-protection, a t-butyl ester for the β-carboxyl of aspartic acid, and a p-alkoxybenzyl alcohol ester for the anchoring bond to the resin support. The nonapeptide showed growth hormone activity in the hypophysectomized rat tibia epiphyseal width test, but was inactive in the hypophysectomized rat body weight gain test.International journal of peptide and protein research 02/1974; 6(2):103-9.
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ABSTRACT: Growth Hormone (GH) gene is a member of gene family through the evolutionary process from a small common ancestral gene by a series of gene duplications. The role of the GH in growth and performance controls has been extensively studied in human, mice and livestock. Many researchers have considered GH as a strong candidate gene for evaluation of genetic polymorphisms that could be associated with economic traits in cattle. We report here a novel missense mutation within the exon 5 of the bovine Growth Hormone (bGH) gene. We could amplified 522 bp fragments from eight unrelated Hanwoo cattle by PCR, then, subsequently cloned and sequenced. An Msp I RFLP corresponding to a C to T transition was observed at position 2258 nt. From this result, we could predict a missense mutation (Arg to Trp) at codon 166 in a highly conserved region among many mammals. Codominant Mendelian segregation of the two alleles, Msp I (+) and Msp I (-), was observed in two full-sib F2 families (n = 32, African taurine Bos taurus African zebu Bos indicus) and eight half-sib Hanwoo families. For the availability of genetic marker, we have performed PCR-RFLP with a large number of individual animals from 15 different cattle breeds (European and Asian taurines, and African indicines). Consideration of breed frequencies of Msp I (-) allele in relation to breed type and their geographic origins, shows higher frequencies in humped breeds or Asian cattle breeds than in humpless or European breeds. This result indicates that the missense mutation can be contributed the functional significance such as the signal transduction through the receptor binding, also may be used as a marker for selection of the economic traits in Hanwoo.Journal of Animal Science and Technology. 01/2003; 45(1).
V olume 16, number 3
THE AMINO ACID SEQUENCE OF BOVINE GROWTH HORMONE
J.A. SANTOM& J.M. DELLACHA, A.C. PALADINI, C.E.M. WOLFENSTEIN
C. PERA, E. POSKUS, S.T. DAURAT, M.J. BISCOGLIO
Z.M.M. DE SE& and A.V.F. DE SANGUESA
Fact&ad de Farmacia y Bioquimica, Departamento
para el Estudio de las Hormonas Hipofisarias, Junin 956, Buenos Aires, Argentina
de Quimica Biolbgica y Centro
Received 4 June 1971
Bovine growth hormone is one of the most
studied growth hormones but its complete pri-
mary structure has not been established until now.
For several years it was accepted that the mole-
cular weight of bovine, as well as porcine and ovine
growth hormones was approximately
this value being clearly different from that of the
hormones of human and monkey origin that were
only half as big [2-41.
true molecular weights of the hormones obtained
from all the abovementioned
and close to 21,000 [S-8] . This value accounts
for the number of disulphide bridges and tryptic
peptides in bovine growth hormone
This paper reports the amino acid sequence of
the bovine growth hormone molecule.
46,000 [ 1 ] ,
have established that the
species are similar
Bovine growth hormone of high purity was pre-
pared by the method of Dellacha and Sonenberg
 . Amino acid analyses and oxidation of the
protein were done as described previously
The oxidized product was submitted
with trypsin, chymotrypsin
or pepsin. The tryptic
digestion was carried out as indicated by SantomC.
Wolfenstein and Paladini [ 1 l] ; the chymotryptic
digestion was done at pH 8, in a pH-stat during 6 hr
at XI”, with a weight ratio of enzyme to hormone
of 1:66. The digestion with pepsin was performed
in 5% formic acid at 37” during 16 hr, with a weight
ratio of enzyme to hormone of 1: 10.
The soluble fraction of the various digests was
fractionated on ion-exchange
procedures similar to those described by Margoliash
and Smith [ 121. Some of these fractions required
further purification by high-voltage electrophoresis
[ 131 on paper, using volatile buffers of pH 6.45
and 2.0 and by two-dimensional
graphy in the following solvent systems: l-butanol-
formic acid-water (75: 15: 10, v/v) and l-butanol-
2.8% ammonium hydroxide-ethanol
When the size of a polypeptide
sidered too big for direct sequencing it was sub-
mitted to partial hydrolysis in 12 M hydrochloric
acid at 40” during 21 hr. The resulting mixture was
resolved by paper chromatography
To avoid the insoluble residue arising in the tryp-
tic digestion of the oxidized hormone this fragmen-
tation was alternatively carried out on the reduced,
[ 141 and maleinized protein
The amino acid sequence in the purified peptide
fragments was determined
resin columns using
(50:20: 15, v/v).
fragment was con-
as indicated be-
from the N-terminal
Volume 16, number 3 FEBS LETTERS
Pr o- Thr - Gl y- Lys- Asn- G1u- Al a~Gl n- Gl u~Lys- Ser - Asp- Leu- Gl u- Leu~Leu- Ar g- I l e- Ser - Leu-
Phe- Gl y- Gl u- Al a- Ser - Cys- Al a- Phe- OH
Fi g. 1. Amino acid sequence of bovine growth hormone. Numbers indicate the position of the amino acid residues from the ammo
terminu_s; t indicates points of tryptic attack; $ indicates points of chymotryptic
,I : indicates points of tryptic attack on the maleinized protein; 0 indicates points of attack by cyanogen bromide.
attack. , : indicates points of peptic attack;
end by the stepwise phenylisothiocyanate
Edman [ 161. The procedures applied were similar to
those described by Konigsberg and Hill [ 171 or by
Elzinga [ 181. In certain cases the sequence was estab-
lished by applying the dansyl method of Gray and
Hartley [ 191 to the residual peptide. The C-terminal
sequence of the same fragments was studied by
digestions with carboxypeptidases
A and B [ 141.
3. Results and discussion
The amino acid sequence indicated in fig. 1 was
confirmed by the amino acid composition of the
fragments obtained treating the hormone with
cyanogen bromide, as previously described [ 14,201.
The location of one disulphide bridge was thus es-
tablished to occur between residues 53 and 161.
The disulphide bridge near the C-terminal end of
the molecule had already been identified
single tryptophan residue is in position 86. The
three histidine residues are in positions
Thirty percent of the molecules have valine re-
placing leucine 124  . This finding substantiates
the microheterogeneity suggested by the amino
acid composition of a tryptic peptide from bovine
growth hormone reported by Fellows and Rogol .
[ 141. The
Volume 16, number 3
The sequence found in the first 2 1 amino acids
from the N-terminal end of the molecule coincides
with the results obtained by Fellows, Rogol and
Wallis  has obtained evidence of the exis-
tence of two forms of bovine growth hormone,
presumably differing only by the presence or ab-
sence of the alanine in position 1. We have con-
firmed these findings in the present investigation.
Bovine growth hormone has about 61% of its
amino acid residues identical and in the same
sequence as those in human growth hormone,
when the comparison is made with the structure
reported by Li, Dixon and Liu  as revised by
Niall  .
The authors wish to thank Dr. R.E. Fellows
for helpful discussion and exchange of information
about the amino acid sequence on the N-terminal
end of bovine growth hormone; Mr. H.N. Fernln-
dez for his contribution to the elucidation
tions 137- 138 and 144- 145 in the chain; Miss
Dora M. Beatti for her skilled and devoted techni-
cal assistance and the able help of Nbtor L. Astorri.
This work was supported in part by grants from
the Consejo National de Investigaciones
y Tecnicas de la Republica Argentina.
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