Resolution and biological properties of three N-terminal analogues of recombinant human interleukin-1 beta.
Recombinant human interleukin-1 beta (rhuIL-1 beta) was purified to apparent homogeneity using standard procedures. The protein was characterized by N-terminal sequence analysis and found to consist of three forms: Met (20%), Ala (75%), and des-Ala (5%). Utilizing TSK SP 5PW HPLC, each form was resolved and analyzed by 2-D acrylamide gel electrophoresis. The analogues were identical in molecular mass (17,500 d) but differed in pI (Met, pI = 6.7; Ala, pI = 6.8; des-Ala, pI = 6.8). Bioactivity measurements, using the C3H/HeJ thymocyte proliferation assay, showed that native Ala rhuIL-1 beta was 300-400% more active than des-Ala rhuIL-1 beta, and was 400-600% more active than Met rhuIL-1 beta protein.
Yem
A W
AW
Biopolymer Chemistry, Upjohn Company, Kalamazoo, MI 49001.
Richard
K A
KA
Staite
N D
ND
Deibel
M R
MR
D000818:Animals; D001681:Biological Assay; D002455:Cell Division / Q000187:drug effects; D006801:Humans; D007375:Interleukin-1 / Q000302:isolation & purification* / Q000502:physiology; D053583:Interleukin-1beta; D051379:Mice; D008809:Mice, Inbred C3H; D010446:Peptide Fragments / Q000302:isolation & purification* / Q000502:physiology; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000502:physiology; D013950:Thymus Gland / Q000166:cytology / Q000187:drug effects
D007375:Interleukin-1; D053583:Interleukin-1beta; D010446:Peptide Fragments; D011994:Recombinant Proteins; C052994:interleukin-1beta (163-171)