PMID- 9518454
OWN - NLM
STAT- MEDLINE
DCOM- 19980526
LR  - 20131121
IS  - 1046-5928 (Print)
IS  - 1046-5928 (Linking)
VI  - 12
IP  - 2
DP  - 1998 Mar
TI  - Expression of human monocyte chemoattractant protein-1 in the yeast Pichia 
      pastoris.
PG  - 145-50
AB  - The human monocyte chemoattractant protein-1 (MCP-1) was expressed at high levels 
      in Pichia pastoris with the alcohol oxidase promoter. It was secreted from the 
      yeast when either its natural signal sequence or the Saccharomyces cerevisiae 
      alpha-factor signal peptide was used. SDS-PAGE and Western blot revealed two 
      immunoreactive MCP-1 species at 15 and 8.5 kDa designated MCP-1H and MCP-1L, 
      respectively; both were purified by cation-exchange chromatography. MCP-1H could 
      be converted to MCP-1L by treatment with peptide N-glycosidase F, showing that 
      the former is an N-glycosylated form of the latter. Laser desorption mass 
      spectrometry showed that MCP-1L actually consisted of a mixture of three 
      polypeptides of 8449, 8614, and 8780 Da and MCP-1H showed a broad peak at 11,134 
      Da. N-terminal peptide sequencing indicated that nearly half of MCP-1L lacked the 
      two N-terminal amino acids found in the native protein. Both MCP-1H and MCP-1L 
      could induce monocyte migration and calcium influx in THP-1 monocytic leukemia 
      cells, although these activities were about 10- to 100-fold lower than those of 
      MCP-1 produced in insect cells.
FAU - Beall, C J
AU  - Beall CJ
AD  - Neurobiotechnology Center, Ohio State University, Columbus, Ohio 43210, USA.
FAU - Breckenridge, S M
AU  - Breckenridge SM
FAU - Chakravarty, L
AU  - Chakravarty L
FAU - Kolattukudy, P E
AU  - Kolattukudy PE
LA  - eng
GR  - HL 48916/HL/NHLBI NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Protein Expr Purif
JT  - Protein expression and purification
JID - 9101496
RN  - 0 (Chemokine CCL2)
RN  - 0 (Immune Sera)
RN  - 0 (Protein Sorting Signals)
RN  - 0 (Recombinant Proteins)
RN  - EC 3.5.- (Amidohydrolases)
RN  - EC 3.5.1.52 (Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Amidohydrolases/metabolism
MH  - Amino Acid Sequence
MH  - Animals
MH  - Biological Assay
MH  - Blotting, Western
MH  - Calcium/metabolism
MH  - Chemokine CCL2/*biosynthesis/genetics/metabolism
MH  - Dose-Response Relationship, Drug
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Gene Expression Regulation, Fungal
MH  - Glycosylation
MH  - Humans
MH  - Immune Sera/immunology
MH  - Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
MH  - Pichia/genetics/*metabolism
MH  - Protein Sorting Signals/chemistry/genetics
MH  - Rabbits
MH  - Recombinant Proteins/biosynthesis/genetics/metabolism
MH  - Tumor Cells, Cultured
EDAT- 1998/03/31 00:00
MHDA- 1998/03/31 00:01
CRDT- 1998/03/31 00:00
PHST- 1998/03/31 00:00 [pubmed]
PHST- 1998/03/31 00:01 [medline]
PHST- 1998/03/31 00:00 [entrez]
AID - S1046-5928(97)90820-2 [pii]
AID - 10.1006/prep.1997.0820 [doi]
PST - ppublish
SO  - Protein Expr Purif. 1998 Mar;12(2):145-50. doi: 10.1006/prep.1997.0820.