Cuttlefish sperm protamines

Martin-Ponthieu, Annie; Wouters-Tyrou, Danièle; Bélaïche, Denise; Sáutière, Pierre; Schindler, Patrick; Dorsselaer, Alain Van

doi:10.1111/j.1432-1033.1991.tb15744.x

Cited by 21 publications

(14 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The minor one (C-1') would be : R6-S-R5-S-R4-S-R-S-P-Y, but its fragmentation is not visible in the FAB mass spectrum. This was later confirmed by the direct sequencing of separated variants Spl and Sp2 [16] whch were both detected by ES-MS. Table 2). From the intense fragmentation pattern observed in FAB-MS (Fig.…”

Section: Chvmotrvptic Peptides C-i C-2b C-2h' C-2a and C-3mentioning

confidence: 70%

“…The sequence of the two variants, corresponding to the two major series of multiply charged ions C and A, was established as being Spl (C) and Sp2 (A) described in the accompanying paper [16].…”

Section: Protamines Sp From Cuttlefishmentioning

confidence: 99%

“…Amino acid analysis and automated sequencing data yielded several possible structures for these five chymotryptic peptides (Table 1) [16]. To eliminate ambiguities, peptides C-1, C-2b, C-2b' and C-3 were submitted to FAB-MS and to ' "Cf PD-MS. Peptide C-3 was also subjected to ES-MS. C2a was only analysed by FAB-MS.…”

Section: Chvmotrvptic Peptides C-i C-2b C-2h' C-2a and C-3mentioning

confidence: 99%

See 2 more Smart Citations

Cuttlefish sperm protamines

Schindler

Bitsch

Klarskov³

et al. 1991

European Journal of Biochemistry

Self Cite

View full text Add to dashboard Cite

The sequence of very basic proteins such as protamines (more than 50% arginines) and related peptides has been determined using mass spectrometry in conjunction with Edman degradation. The capabilities of three mass spectrometric (MS) techniques [fast-atom-bombardment (FAB), ' "Cf plasma desorption ("'CFPD) and electrospray (ES)] have been evaluated on stallion protamine 1, cuttlefish protamine, and the corresponding cleavage peptides. In contrast to FAB-MS and "' Cf PD-MS, ES-MS made possible an easy determination of the molecular mass of the intact protamines ( NN 8 kDa). With ES-MS about 0.2 nmol was sufficient to yield a mass measurement with an accuracy of 0.05%. On peptides smaller than 3500 Da, both FAB-MS and ' "Cf PD-MS allowed mass measurements with an accuracy of 0.1%. 252Cf PD-MS appeared more sensitive than FAB-MS by ~ about a factor of 10. FAB-MS is nevertheless particularly interesting since in most cases it produced spectra with intense A-type fragmentation ions which provided reliable primary structure information. Protamines are highly basic proteins, associated with DNA in sperm nuclei. They are characterized by a very high arginine content (more than 50%). Purifications of protamines and cleavage peptides from enzymic digestions by ionexchange chromatography or reverse-phase HPLC are difficult because of close similarities in the amino acid composition. Primary structure elucidation of such proteins is also particularly difficult when using conventional methods of protein analysis. On purified protamines and cleavage peptides, amino acid analysis yields poor information because of the disproportion between the arginines and the other residues, especially hydrophobic residues. Moreover, serine and threonine, which often appear in the amino acid composition of such polypeptides, are partially destroyed during acid hydrolysis and cannot constitute a reference for the quantification. Primary structure elucidation of such polypeptides by automated Edman degradation is often ambiguous because the repetitive yield of the phenylthiohydantoin of arginine decreases rapidly when arginines are consecutive. New methods for purity evaluation, as well as for sequence determination, would therefore be very useful in the field of protamines.

show abstract

Section: Chvmotrvptic Peptides C-i C-2b C-2h' C-2a and C-3mentioning

confidence: 70%

Section: Protamines Sp From Cuttlefishmentioning

confidence: 99%

Section: Chvmotrvptic Peptides C-i C-2b C-2h' C-2a and C-3mentioning

confidence: 99%

See 1 more Smart Citation

Cuttlefish sperm protamines

Schindler

Bitsch

Klarskov³

et al. 1991

European Journal of Biochemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…It is. an interesting case of convergent evolution, since such an alternating region is absent in the protamines of fish [37] and in the cephalopods, another group of molluscs [38].…”

Section: Structural Comparisonmentioning

confidence: 99%

Helical structure of basic proteins from spermatozoa

Verdaguer

Perello

Palau

et al. 1993

European Journal of Biochemistry

View full text Add to dashboard Cite

We describe structural studies carried out with some basic proteins found in association with DNA in the spermatozoa of molluscs and echinoderms. We have studied proteins related to histone H1 as well as protamines. Structural prediction methods show that these proteins have a strong helical potential and contain several turns, mainly of the SPKK type. No p structures were found.Strong structural similarities have been detected between distantly related species. The presence of helical regions is confirmed by circular dichroism in trifluoroethanol solution. The influence of the SPKK turns is also evident in the CD spectra. In proteins which contain a high percentage of arginine we conclude that conventional prediction methods should be modified in order to allow for a higher helical potential for this amino acid residue. Synthetic peptides with a sequence present in the C-terminal region of histone H1 have also been studied. It was found that octapeptides may only acquire a small amount of structure, whereas hexadecapeptides are 50-60% helical. These studies strongly suggest that both protamines and proteins related to the C-terminal part of histone H1 interact with DNA mainly in the a-helical conformation.The basic proteins associated with DNA in the nuclei of spermatozoa show a wide range of variation in number of components, amino acid composition and size, as reviewed elsewhere [l]. In some cases a complete set of somatic histones may be found, whereas in other cases histones have been completely substituted by one or several arginine-rich proteins (protamines). In some species intermediate proteins are found. In this paper we analyze the secondary structure of several sperm proteins which appear to be related to histone H1 and compare their structure with protamines.It is known [2] that histone H1 is involved in the formation of the higher-order structure of chromatin fibers. It has also been shown that this histone has three clearly defined structural domains [3]. Later it was found [4-61 that the Cterminal domain of this histone, which is rich in Lys, Ala and Pro, has regions with a high helical potential, in particular in histone H1 from sea urchin spermatozoa. Models have been proposed [7-91 in order to explain how these helical regions might stabilize the higher-order structure of DNA.Recently we have shown [lo] that chromatin fibers may be formed in mollusc spermatids and spermatozoa in the presence of protamines and H1-related proteins, but with a very low content of core histones. In organisms in which only arginine-rich protamines are present, no chromatin fibres are found, but instead the nucleus is very compact [ l l ] and the nucleoprotamine complex is a highly ordered system of parallel DNA molecules [12]. One of us has recently sug- gested that protamines may also stabilize this complex in a helical conformation [7, 81. In view of this situation and due to the fact that the sequence of some of these proteins has been recently determined [13, 141, we decided to study the structural properties...

show abstract

“…2, lane 7 and Fig. 3, panel 4) (36). It is important to note that the arginine content of PL-I can be as high as 35 mol % in some bivalve organisms (37).…”

mentioning

confidence: 99%