1H and 15N chemical shift assignments for HP-RNase

El-Joubary, A., Bruix, M., Santoro, J., Cafaro, V., Scognamiglio, R.,
Di Donato, A., D'Alessio, G., Kover, K.E., Batta, G., Szilagyi, L, and
Rico, M., "1H and 15N Sequential Assignment and Solution Secondary Structure
of 15N Labelled Human Pancreatic Ribonuclease," J. Biomol. NMR 15, 265-266 (1999).

1H and 15N sequential assignment and solution secondary structure of 15N labelled human pancreatic ribonuclease.

The primary structure of human (Homo sapiens) pancreatic ribonuclease has been
determined by automatic sequencing of the native protein and by analysis of
peptides obtained by cleavage with proteolytic enzymes, cyanogen bromide, and
hydroxylamine. The following sequence was deduced: (sequence in text). Human
pancreatic ribonuclease differs at 37 positions from bovine pancreatic
ribonuclease. In addition the human enzyme has three more residues at the
C-terminus. About half of the enzyme molecules contain carbohydrate attached to
the sequence Asn-Met-Thr (34-36). Two other Asn-X-Ser/Thr sequences are
carbohydrate free. Human pancreatic ribonuclease contains many positively
charged residues, especially near the N-terminus, while negatively charged
residues are more concentrated near the C-terminus.

Beintema, J.J., Wietzes, P. Weickmann, J.L. and Glitz, D.G.
Anal. Biochem. (1984) 136, 48-64

The amino acid sequence of human pancreatic ribonuclease.

A synthetic cDNA coding for human pancreatic RNase, equipped with a secretion
signal sequence, was cloned and stably expressed in Chinese hamster ovary
cells. The recombinant RNase, secreted into the culture medium, was purified
and characterized. It was found to be indistinguishable, by structural and
catalytic parameters, from the enzyme isolated from human pancreas.
Furthermore, the glycosylated forms were separated from the non-glycosylated
form. Up until now, human RNases have been isolated only in small amounts from
autopic specimens. This has hindered the exploitation of a human RNase for the
construction of immunotolerated immunotoxins. On the other hand, the
availability of an effective system for the expression of a human RNase may
render feasible the transfer, by protein engineering, of the interesting
pharmacological actions of non-human RNase [1993 Trends Cell Biol. 3, 106-109]
to an immunotolerated, human RNase.

Russo, N., de NIgris, M., Ciardello, A., Di Donato, A and D'Alessio, G.
FEBS Lett. (1993) 333, 233-237.
Anal. Biochem. (1984) 136, 48-64

Expression in mammalian cells, purification and characterization of recombinant human pancreatic ribonuclease.

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.

Piccoli, R., Di Gaetano, S., De Lorenzo, C., Grauso, M., Monaco, C., Saplleti
Cernia, D., Lacetti, P. Cinatl, J., Matousek, J. D'Alessio, G.
Proc. Natl. Academ. Sci. USA (1999) in press

Proceedings of the National Academy of Sciences of the United States of America
A dimeric mutant of human pancreatic ribonuclease with selective cytotoxicity toward malignant cells.

Youle, R.J. and D'Alessio, G.
Ribonucleases. Structure and Functions (D'Alessio, G. & Riordan, F.J. Eds)
(1997)
pp491-514, Academic Press, New York, NY.

antitumor potential-structure relationship

The recombinant protein obtained has a Met residue at the position preceding Lys 1

Ribonuclease 1 Des1-7 Crystal Structure At 1.9a

X-Ray Structure Of Human Ribonuclease Inhibitor Complexed With Ribonuclease I

Mutant Human Ribonuclease 1 (r31l, R32l)

Mutant Human Ribonuclease 1 (Q28l, R31l, R32l)

Solution Structure Of Human Pancreatic Ribonuclease

Ensemble Refinement Of The Protein Crystal Structure Of Human Ribonuclease Inhibitor Complexed With Ribonuclease I

BerH2-scFv-hpRNase precursor [Drosophila melanogaster]

Ribonuclease, RNase A family, 1 (pancreatic) [Homo sapiens]

pancreatic ribonuclease [Pan troglodytes]

pancreatic ribonuclease [Gorilla gorilla]

ribonuclease pancreatic precursor [Pan troglodytes]

ribonuclease pancreatic precursor [Homo sapiens]

RecName: Full=Ribonuclease pancreatic; AltName: Full=HP-RNase; AltName: Full=RIB-1; AltName: Full=RNase UpI-1; AltName: Full=Ri

RecName: Full=Ribonuclease pancreatic; AltName: Full=RNase 1; AltName: Full=RNase A; Flags: Precursor

TPA: ribonuclease A C1 [Pan troglodytes]

TPA: ribonuclease A C1 [Nomascus leucogenys]