Chemical method for selective modification of the N- and/or C-te

Details Chemical method for selective modification of the N- and/or C-te Chemical method for selective modification of the N- and/or C-te

1994-08-23

1997-06-03

Patterson, Jr., Charles L.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

4351723, C12P 2100

Patent

active

056353715

ABSTRACT:
The invention provides for a chemical method for preparing a recombinant single copy polypeptide or a portion thereof with a modified terminal amino acid .alpha.-carbon reactive group selected from the group consisting of N-terminal .alpha.-amine, C-terminal .alpha.-carboxyl, and a combination thereof. The steps of the method involve forming the recombinant single copy polypeptide or a portion thereof so that the single copy polypeptide is protected with one or more biologically added protecting groups at the N-terminal .alpha.-amine, C-terminal .alpha.-carboxyl. The recombinant single copy polypeptide can then be reacted with up to three chemical protecting agents to selectively protect reactive side chain groups and thereby prevent side chain groups from being modified. The recombinant single copy polypeptide can be cleaved with at least one cleavage reagent specific for the biological protecting group to form an unprotected terminal amino acid .alpha.-carbon reactive group. The unprotected terminal amino acid .alpha.-carbon reactive group is modified with at least one chemical modifying agent. The side chain protected terminally modified single copy polypeptide is then deprotected at the side chain groups to form a terminally modified recombinant single copy polypeptide. The number and sequence of steps in the method can be varied to achieve selective modification at the N- and/or C-terminal amino acid of a recombinantly produced polypeptide.

REFERENCES:
patent: 4431739 (1984-02-01), Riggs
patent: 4801536 (1989-01-01), Stahl et al.
patent: 4810777 (1989-03-01), Zasloff
patent: 5004686 (1991-04-01), Cohen et al.
patent: 5045531 (1991-09-01), Berkowitz et al.
patent: 5137872 (1992-08-01), Seely et al.
patent: 5166321 (1992-11-01), Lai et al.
Schellenberger et al., "Proteinase-catalyzed conversion of a substance P-precursor peptide," Int. J. Peptide Protein Res., 39, 1992, pp. 472-476.
Schellenberger et al., "Peptide production by a combination of gene expression, chemical syntheses, and protease-catalyzed conversion," Int. J. Peptide Protein Res., 41, 1993, pp. 326-332.
Morihara, "Using proteases in peptide syntheses," TibTech, vol. 5, Jun. 1987, pp. 164-170.
Carles et al., "C-terminal labelling of .beta.-casein," FEBS Letters, vol. 212, No. 1, Feb. 1987, pp. 163-167.
Bongers et al., "Comparison of enzymatic semisyntheses of peptide amides: human growth hormone releasing factor and analogs," Biomed. Biochim. Acta, Acta, 50 1991, pp. S157-S162.
Bongers et al., "Semisynthesis of human growth hormone-releasing factor by trypsin catalyzed coupling of leucine amide to a C-terminal acid precursor," Int. J. Peptide Protein Res., 40, 1992, pp. 268-273.
Fruton, "Proteinase-catalyzed synthesis of peptide bonds," Advances in Enzymology, vol. 53, A. Meister, Ed., 1982, pp. 239-306.
Cha et al., Biochemical Pharm., 30, 1981, pp. 1507-1515.
Kempe et al., Biotechnology, 4, 1992, pp. 565-568.
Shen, Proc. Natl. Acad. Sci. USA, 81, 1984, pp. 4627-4631.
Science, vol. 177, 18 Aug. 1972, Lancaster, PA, US, pp. 623-626, M.A. Ruttenberg, "Human Insulin: Facile Synthesis by Modification of Porcine Insulin".
Hoppe-Seyler's Zeitschrift Fur Physiologischen Chemie, vol. 357, Jun. 1976, Berlin D., pp. 759-767, R. Obermeir and R. Geiger, "A New Semisynthesis of Human Insulin".
Agarwal et al., Biochemical Pharm., 26, 354-367 (1977).
R. Barker, Methods Enzymol., 34, 317-328 (1974).
R. Bateman et al., J. Biol. Chem., 260, pp. 9088-9091 (1985).
S. Beaucage et al., Tetra. Letters, 221:859 (1981).
G.S. Bethell et al., J. Biol. Chem., 254, 2572-2574 (1979).
Cha et al., Biochemical Pharm., 24, 2187-2197 (1975).
Degrado et al., J. of Cellular Biochem., 29, 83-93 (1985).
J. Engels, Protein Engineering, 1:195-199 (1987).
Hanada et al., J. Biol. Chem., 263:7181 (1988).
J. Johansen, Carlsberg Res. Commun., 41, 73 (1976).
K. Itakura et al., Science, 198:1056 (1977).
Kirk-Othmer Encyclopedia of Chemical Technology, 4th Edition, vol. 12, pp. 603-617 (1991).
B. Lewin, Genes, 4th Edition, pp. 606-608, Oxford Press, New York, NY (1990).
Li et al., PNAS, 80, 2216 (1983).
S. Marcus, Methods Enzymol., 34, 377-385 (1974).
I. Matsumoto, Methods Enzymol., 34, 329-341 (1974).
A. Matsuura et al., Methods Enzymol., 34, 303-4 (1974).
B. Nilsson et al., EMBO Journal, 4:1075 (1985).
J. Pierce et al., Biochem., 19:934-942 (1980).
Ray et al., 201st ACS National Meeting, Atlanta, GA, Abstract Papers Am. Chem. Soc. 201, Abstract Biot. 48 (1991).
W. Scouten, Methods Enzymol., 34, 288-294 (1974).
Spindel et al., PNAS, 81, 5699 (1984).
Sutton et al., BBRC, 134, 386-392 (1986).
Tajima et al., J. Fermentation & Bioengineering, 72:362 (1991).
Taylor, P.W. et al., Biochemistry, 9, 2638 (1970).
Williams et al., Biochemical Pharm., 29, 589-595 (1980).
Yoo et al., J. Biol. Chem., 264:17078 (1989).
Bodanszky (1988) Peptide Chemistry, pp. 1-9 and 55-103.
Kempe et al. Gene vol. 39 pp. 239-245 (1985).
Bullesbach et al. Biochem. vol. 25 pp. 5998-6004 (1986).
Lehninger, Biochemistry, "The Molecular Basis of Cell Structure and Function", 1975, pp. 202-206.

Affiliated with

Also associated with

Rating

Chemical method for selective modification of the N- and/or C-te does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Chemical method for selective modification of the N- and/or C-te, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Chemical method for selective modification of the N- and/or C-te will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-390685