Complete amino acid sequence of boar protamine

Tobita, Tohru; Tsutsumi, Hiroyuki; Kato, Akemi; Suzuki, Hiromasa; Nomoto, Minoru; Nakano, Minoru; Ando, Toshio

doi:10.1016/0167-4838(83)90083-3

Cited by 67 publications

(31 citation statements)

References 23 publications

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“…Also the amino acid compositions (table 1) were virtually identical and typical for protamines, i.e. very rich in arginine and half-cystine, but devoid of alanine, which otherwise is part of the characteristic N-terminus of mammalian type 1 protamines [5,[7][8][9][10][11][12][13][14]. The composition is similar to those published earlier [ 17,18].…”

Section: Resultssupporting

confidence: 72%

“…The alignment demonstrates that strongly basic arginine clusters, which presumably bind to the acidic DNA, tyrosines, which may act as DNA intercalators, and most half-cystines, which disulfide-crosslink sperm chromatin, all are evolutionarily conserved. The presence of valine instead of alanine at the Nterminus is a surprise, as the constancy of Nterminal alanine in all previously sequenced mammalian type 1 protamines [5,[7][8][9][10][11][12][13][14] seemed to indicate its functional importance. In fact, many non-mammalian arginine-rich protamines also possess an N-terminal alanine, the N-terminal sequences being reminiscent of the mammalian counterparts, i.e.…”

Section: Discussionmentioning

confidence: 99%

“…These two types show no convincing structural homology. However, human and mouse type 1 protamines are homologous with the single (or main) protamines of boar [10], ram [11], bull [12] and stallion [13,14]. It could be expected that proteins forming the corresponding, compact complexes with DNA show a high degree of mutual resemblance and are evolutionary conserved, and indeed the six protamine sequences contain similar, highly basic arginine clusters, which bind to the acidic DNA.…”

Section: Introductionmentioning

confidence: 99%

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Primary structure of rabbit sperm protamine, the first protamine of its type with an aberrant N‐terminal

Ammer

Henschen

1988

FEBS Letters

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Rabbit protamine was extracted from S-(pyridylethylated) sperm cell nuclei with hydrochloric acid and then isolated by reversed-phase HPLC. The primary structure was determined by amino acid sequence analysis of the total protein and of fragments obtained by digestion with endoproteinase Lys-C and thermolysin. The protamine contains 49 amino acid residues and is clearly homologous with mammalian type I protamines, 47% of the positions being invariant. Surprisingly, rabbit protamine possesses an N-terminal valine residue, whereas all mammalian and several non-mammalian protamine sequences of this type start with alanine, the N-terminal region being remarkably conserved during evolution.

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Section: Resultssupporting

confidence: 72%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Primary structure of rabbit sperm protamine, the first protamine of its type with an aberrant N‐terminal

Ammer

Henschen

1988

FEBS Letters

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“…Protamine P2a is approximately 50% homologous with human protamine PI. The amino acid sequence of P2a is: In a previous paper [I 31 we reported the complete amino acid sequence of human protamine PI whch was found to be quite homologous to the single protamine in bull [I], ram [2] and boar [3] as well as to mouse protamine 1 [14]. In this paper we present the complete amino acid sequences of two forms of the second major human protamine, P2.…”

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confidence: 99%

Human sperm protamines. Amino-acid sequences of two forms of protamine P2

1986

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Human protamine P2 was purified to homogeneity by solubilizing whole spermatozoa in guanidinium . HC1 containing 2-mercaptoethanol, alkylating the resulting protamine thiols with vinylpyridine, removing acid-insoluble material by acid dialysis and using CM-cellulose chromatography to remove non-protamine basic proteins and separate protamines P1 and P2. The P2 preparation contained two components, P2a and P2b, which were sequenced completely without being separated. The peptides obtained from thermolysin and endoproteinase Lys-C digestions were purified by reverse-phase high-pressure liquid chromatography and sequenced using a gasphase sequencer. P2a contains 57 amino acids and has a relative molecular mass of 7636 while P2b contains 54 amino acids, which are identical to residues 4-57 of P2a, and has a relative molecular mass of 7242. Protamine P2a is approximately 50% homologous with human protamine PI. The amino acid sequence of P2a is: In a previous paper [I 31 we reported the complete amino acid sequence of human protamine PI whch was found to be quite homologous to the single protamine in bull [I], ram [2] and boar [3] as well as to mouse protamine 1 [14]. In this paper we present the complete amino acid sequences of two forms of the second major human protamine, P2. MATERIALS AND METHODS MaterialsThe materials used have been described previously [13] Methods Two purification methods were used to isolate human protamine P2. The first method was the modified procedure of Kolk and Samuel [9] which was used to obtain pure protamine PI [13]. Whole spermatozoa from 8 -10 fertile donors were washed, dissolved in 6M guanidinium . HC1 containing 2-mercaptoethanol and alkylated with vinylpyridine. The resulting solution was dialyzed against 2% HCl and the dialysate supernatant was fractionated on a Bio-Rex 70 ionexchange column. The second method was the same as the first except that CM-cellulose [I51 was used in place of the Bio-Rex 70 in the ion-exchange step. CM-cellulose (CM-52, Whatman) was equilibrated with 5% guanidinium . HCI in 50 mM lithium acetate buffer at pH 5.0 and packed into a 1 .0-cm-diameter column on top of a 0.5-cm bed of Sephadex G-10 to a height of 8 cm. The acid dialysate supernatant was mixed with 2.5 g guanidinium . HC1, diluted to 50 ml with lithium acetate buffer and run into the column under gravity at room temperature. Only the very basic protamines bound to the CM-cellulose and they were eluted with a linear gradient of 5 -15% guanidinium . HC1 in lithium acetate buffer in a total volume of 200ml. The fractions under each 230-nm absorbance peak were pooled, exhaustively dialysed against 1% acetic acid in Spectrapore 3 tubing, dried down in a vacuum centrifuge, redissolved in 0.2 M acetic acid and stored at -20 "C. Amino acid analysis, automated protein sequencing and reverse-phase peptide HPLC were performed as described previously [13].Protamine P2 was digested with thermolysin by dissolving 18 nmol in 0.2 ml0.5% ammonium bicarbonate, adding 2 pg thermolysin and incubating the solu...

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“…79:177a, 1978), rats (20,21), bulls (10,32,36), boars (36,43), rams (29,36,41), stallions (36), guinea pigs (8,9), rabbits (8,9), and humans (2,12,25,33,39). Complete sequence analysis of the P1 protamines from bulls (10,32), boars (43), rams (41), and humans (2,33) and the predicted amino acid sequence derived from a cDNA sequence for mouse protamine 1, (mPl) (24) have revealed identical lengths of 50 amino acids and strong sequence homologies among these mammalian protamines. P1 protamine is organized into three domains consisting of a central basic core with clusters of arginine and two less basic amino-and carboxyl-terminal regions.…”

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confidence: 99%

Mouse protamine 2 is synthesized as a precursor whereas mouse protamine 1 is not.

Yelick¹,

Balhorn²,

Johnson³

et al. 1987

Mol. Cell. Biol.

147

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The nuclei of mouse spermatozoa contain two protamine variants, mouse protamine 1 (mPl) and mouse protamine 2 (mP2). The amino acid sequence predicted from mPl cDNAs demonstrates that mPl is a 50-amino-acid protein with strong homology to other mammalian P1 protamines. Nucleotide sequence analysis of independently isolated, overlapping cDNA clones indicated that mP2 is initially synthesized as a precursor protein which is subsequently processed into the spermatozoan form of mP2. The existence of the mP2 precursor was confirmed by amino acid composition and sequence analysis of the largest of a set of four basic proteins isolated from late-step spermatids whose synthesis is coincident with that of mPl. The sequence of the first 10 amino acids of this protein, mP2 precursor 1, exactly matches that predicted from the nucleotide sequence of cDNA and genomic mP2 clones. The amino acid composition of isolated mP2 precursor 1 very closely matches that predicted from the mP2 cDNA nucleotide sequence. Sequence analysis of the amino terminus of isolated mature mP2 identified the final processing point within the mP2 precursor. These studies demonstrated that mP2 is synthesized as a precursor containing 106 amino acids which is processed into the mature, 63-amino-acid form found in spermatozoa.Protamines are small, basic, arginine-rich proteins that replace histones in the later stages of spermatogenesis in many vertebrates (N. B. Hecht, in G. Stein and J. Stein, ed., Basic Chromosomal Proteins: Structure, Organization and Regulation ofthe Gene, in press). During spermatogenesis in mammals, the histones are not directly replaced by protamines but by a population of basic proteins that are transiently associated with the spermatid nucleus (14,35; Hecht, in press). These testis-specific proteins are removed from the condensing chromatin at later stages of spermiogenesis and are replaced by protamines, the predominant class of proteins found complexed with DNA in the mature spermatozoon. Mammalian protamines contain the amino acid cysteine, which is believed to function in the stabilization and compaction of the sperm nucleus through disulfide bond cross-linking (7).Protamines have been isolated from a large number of vertebrates including fish (3, 28), domestic fowl (38), mice (6, 27, 40; A. R.

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Complete amino acid sequence of boar protamine

Cited by 67 publications

References 23 publications

Primary structure of rabbit sperm protamine, the first protamine of its type with an aberrant N‐terminal

Primary structure of rabbit sperm protamine, the first protamine of its type with an aberrant N‐terminal

Human sperm protamines. Amino-acid sequences of two forms of protamine P2

Mouse protamine 2 is synthesized as a precursor whereas mouse protamine 1 is not.

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