Isolation and Some Chemical Properties of Aspermatogenic Substance From Bull Seminal Vesicle Fluid

Dostál, J; Matoušek, J.

doi:10.1530/jrf.0.0330263

Cited by 46 publications

(14 citation statements)

References 10 publications

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“…BS-RNase was purified from seminal vesicles as described (15,16). Escherichia coli strain BL21 (DE3) pLysS and expression vector pET22b ϩ were provided by AMS Biotechnology (Milan); E. coli JM 101 strain was from Boehringer Mannheim; restriction enzymes were from Promega.…”

Section: Methodsmentioning

confidence: 99%

“…1). Four of these are located in a helical segment (helix-II, residues 24-34) that is connected to the N-terminal ␣-helix (residues 3-13) by a hinge loop (residues [16][17][18][19][20][21][22]. In BS-RNase, the intersubunit interface is constituted by the helix-II segments of the two subunits (10).…”

Section: Construction and Characterization Of A Dimeric Mutant Of Hp-mentioning

confidence: 99%

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A dimeric mutant of human pancreatic ribonuclease with selective cytotoxicity toward malignant cells

Piccoli

Gaetano

Lorenzo

et al. 1999

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

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Monomeric human pancreatic RNase, devoid of any biological activity other than its RNA degrading ability, was engineered into a dimeric protein with a cytotoxic action on mouse and human tumor cells, but lacking any appreciable toxicity on mouse and human normal cells. This dimeric variant of human pancreas RNase selectively sensitizes to apoptotic death cells derived from a human thyroid tumor. Because of its selectivity for tumor cells, and because of its human origin, this protein represents a potentially very attractive, novel tool for anticancer therapy.

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Section: Methodsmentioning

confidence: 99%

Section: Construction and Characterization Of A Dimeric Mutant Of Hp-mentioning

confidence: 99%

A dimeric mutant of human pancreatic ribonuclease with selective cytotoxicity toward malignant cells

Piccoli

Gaetano

Lorenzo

et al. 1999

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

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“…Seminal RNase, which represents approximately 2% of the total protein in bovine seminal plasma, has antispermatogenic activity (17), immunosuppressive activity (18)(19)(20), and cytotoxic activity against many transformed cell lines (21,22). Each of these activities is largely absent from pancreatic RNase and the protein that was the most probable recent common ancestor of seminal and pancreatic RNase (23).…”

mentioning

confidence: 99%

Origin of Dimeric Structure in the Ribonuclease Superfamily

et al. 1998

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To enable application of postgenomic evolutionary approaches to understand the divergence of behavior and function in ribonucleases (RNases), the impact of divergent sequence on the divergence of tertiary and quaternary structure is analyzed in bovine pancreatic and seminal ribonucleases, which differ by 23 amino acids. In a crystal, seminal RNase is a homodimer joined by two "antiparallel" intersubunit disulfide bonds between Cys-31 from one subunit and Cys-32' from the other and having composite active sites arising from the "swap" of residues 1-20 from each subunit. Specialized Edman degradation techniques have completed the structural characterization of the dimer in solution, new cross-linking methods have been developed to assess the swap, and sequence determinants of quaternary structure have been explored by protein engineering using the reconstructed evolutionary history of the protein family as a guide. A single Cys at either position 32 (the first to be introduced during the divergent evolution of the family) or 31 converts monomeric RNase A into a dimer. Even with an additional Phe at position 31, another residue introduced early in the seminal lineage, swap is minimal. A hydrophobic contact formed by Leu-28, however, also introduced early in the seminal lineage, increases the amount of "antiparallel" connectivity of the two subunits and facilitates swapping of residues 1-20. Efficient swapping requires addition of a Pro at position 19, a residue also introduced early in the divergent evolution of the seminal RNase gene. Additional cysteines required for dimer formation are found to slow refolding of the protein through formation of incorrect disulfide bonds, suggesting a paradox in the biosynthesis of the protein. Further studies showed that the dimeric form of seminal RNase known in the crystal is not the only form in vivo, where a substantial amount of heterodimer is known. These data complete the acquisition of the background needed to understand the evolution of new structure, behavior, and function in the seminal RNase family of proteins.

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“…Originally isolated by D’Alessio [10], seminal RNase displays antiproliferative activity [11, 12], immunosuppressive activity [13–15], antispermatogenic activity [16], and affinity for anionic glycolipids such as gangliosides and seminolipid (N. Trabesinger‐Rüf and S. A. Benner, unpublished results). Each of these activities essentially absent from pancreatic RNase and from the most recent common ancestor of seminal and pancreatic RNase.…”

mentioning

confidence: 99%

Crystal structure of a hybrid between ribonuclease A and bovine seminal ribonuclease – the basic surface, at 2.0 Å resolution

Vatzaki

Allen

Leonidas

et al. 1999

European Journal of Biochemistry

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A variant of bovine pancreatic ribonuclease A has been prepared with seven amino acid substitutions (Q55K, N62K, A64T, Y76K, S80R, E111G, N113K). These substitutions recreate in RNase A the basic surface found in bovine seminal RNase, a homologue of pancreatic RNase that diverged some 35 million years ago. Substitution of a portion of this basic surface (positions 55, 62, 64, 111 and 113) enhances the immunosuppressive activity of the RNase variant, activity found in native seminal RNase, while substitution of another portion (positions 76 and 80) attenuates the activity. Further, introduction of Gly at position 111 has been shown to increase the catalytic activity of RNase against double‐stranded RNA. The variant and the wild‐type (recombinant) protein were crystallized and their structures determined to a resolution of 2.0 Å. Each of the mutated amino acids is seen in the electron density map. The main change observed in the mutant structure compared with the wild‐type is the region encompassing residues 16–22, where the structure is more disordered. This loop is the region where the polypeptide chain of RNase A is cleaved by subtilisin to form RNase S, and undergoes conformational change to allow residues 1–20 of the RNase to swap between subunits in the covalent seminal RNase dimer.

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Isolation and Some Chemical Properties of Aspermatogenic Substance From Bull Seminal Vesicle Fluid

Cited by 46 publications

References 10 publications

A dimeric mutant of human pancreatic ribonuclease with selective cytotoxicity toward malignant cells

A dimeric mutant of human pancreatic ribonuclease with selective cytotoxicity toward malignant cells

Origin of Dimeric Structure in the Ribonuclease Superfamily

Crystal structure of a hybrid between ribonuclease A and bovine seminal ribonuclease – the basic surface, at 2.0 Å resolution

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