Synthesis and properties of carbonylbis(methionyl)insulin, a proinsulin analogue which is convertible to insulin by cyanogen bromide cleavage

Biochemistry
W D Busse, F H Carpenter

Abstract

The preparation and use of carbonylbis (L-methionine p-nitrophenyl ester) as a reversible cross-linking reagent for insulin are described. The reaction of 1 equiv of reagent with zinc insulin in dimethylformamide in the presence of triethylamine yields as one of the products NalphaA1, NepsilonB29-carbonylbis(methionyl)insulin, (CBM-insulin). The CBM-insulin was characterized by end group analysis and by the products formed on tryptic and chymotryptic cleavage. It possessed 91% of the immunological and 6.5% of the hormonal activity of insulin. Treatment of CBM-insulin with cyanogen bromide (CNBr) in 70% formic acid for 1 h resulted in nearly complete removal of the methionine bridge to yield insulin. A small amount of a side product was removed on DEAE-cellulose at pH 7.2 to give an overall recovery of insulin of 70-80%. Oxidative sulfitolyses of CBM-insulin gave the hexa(S-sulfonate) which was reduced with dithiothreitol to yield reduced CBM-insulin. The latter compound, containing 6 sulfhydryls, exhibited a pH-dependent circular dichroic spectrum. The form at pH 10 exhibited a spectrum typical of random coil which was converted to a form at pH 7.8 which was characterized by a negative extremum at 213 nm. The change in the spec...Continue Reading

References

Mar 1, 1973·Analytical Biochemistry·D Racusen
Aug 1, 1974·Analytical Biochemistry·T PooleW W Fish
Sep 4, 1974·Journal of the American Chemical Society·W D BusseF H Carpenter
Jun 15, 1973·Biochimica Et Biophysica Acta·D Levy
Nov 1, 1973·Biochemical and Biophysical Research Communications·R Geiger, R Obermeier
Feb 1, 1973·Hoppe-Seyler's Zeitschrift für physiologische Chemie·W D Busse, H G Gattner
Jun 1, 1973·Hoppe-Seyler's Zeitschrift für physiologische Chemie·D Brandenburg, A Wollmer
Jun 25, 1971·Nature·T L BlundellM Vijayan
Jun 1, 1972·Hoppe-Seyler's Zeitschrift für physiologische Chemie·D Brandenburg
Jun 1, 1968·Proceedings of the National Academy of Sciences of the United States of America·D F Steiner, J L Clark
Feb 20, 1970·Biochemical and Biophysical Research Communications·R W Boesel, F H Carpenter
Mar 1, 1972·FEBS Letters·D G. Lindsay
May 1, 1959·Archives of Biochemistry and Biophysics·G L ELLMAN
Mar 1, 1965·The Biochemical Journal·R F DOOLITTLE
Sep 1, 1960·The Journal of Clinical Investigation·A E RENOLDM C SHEPS
Feb 1, 1967·Proceedings of the National Academy of Sciences of the United States of America·D F Steiner, P E Oyer

Citations

Jan 1, 1980·Diabetologia·D I DronD Brandenburg
Jan 25, 2013·Journal of the American Chemical Society·Michal Avital-ShmiloviciStephen B H Kent
Oct 20, 2009·Journal of the American Chemical Society·Youhei Sohma, Stephen B H Kent
Feb 5, 2014·Genome Biology and Evolution·José Ignacio ArroyoJuan C Opazo
Aug 18, 2010·Angewandte Chemie·Hans-Jürgen Musiol, Luis Moroder
Apr 1, 1984·Analytical Biochemistry·T W ThannhauserH A Scheraga
May 27, 2017·Angewandte Chemie·Luis Moroder, Hans-Jürgen Musiol
Mar 18, 2020·Future Medicinal Chemistry·Florence M BrunelRichard D DiMarchi
Mar 21, 2020·Chemistry : a European Journal·Mads ØstergaardKnud J Jensen
Feb 1, 1979·International Journal of Peptide and Protein Research·R C Paynovich, F H Carpenter
Aug 1, 1983·International Journal of Peptide and Protein Research·B R Srinivasa, F H Carpenter

Related Concepts

Fat Pad
Metazoa
Cyanogen Bromide
Hydrogen-Ion Concentration
Insulin B Chain
Pedameth
Proinsulin
Infrared Spectrophotometry
Spectrophotometry, Ultraviolet
Structure-Activity Relationship

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