PMID- 1429738
OWN - NLM
STAT- MEDLINE
DCOM- 19921222
LR  - 20231213
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 267
IP  - 33
DP  - 1992 Nov 25
TI  - Pseudomonas exotoxin A-epidermal growth factor (EGF) mutant chimeric protein as 
      an indicator for identifying amino acid residues important in EGF-receptor 
      interaction.
PG  - 24034-40
AB  - Epidermal growth factor (EGF) was fused to the carboxyl end of a modified 
      pseudomonas exotoxin A that has its toxin binding domain deleted. This chimeric 
      toxin designated as PE(delta Ia)-EGF kills A431 cells through the EGF 
      receptor-mediated pathway. In this study, we used a random mutagenesis approach 
      to make point mutations on EGF, followed by replacing the wild type EGF in 
      PE(delta Ia)-EGF with these EGF mutants. We have constructed 14 different 
      PE(delta Ia)-EGFmutants, and examined their EGF receptor binding activity as well 
      as their cytotoxicity to A431 cells. Our results showed that individual mutations 
      of Val19 to Glu and Val34 to Asp in the EGF domain of PE(delta Ia)-EGFmutants 
      resulted in an increase in the binding affinity to EGF receptor and cytotoxicity 
      to A431 cells. On the other hand, individual mutations of His16 to Asp and Gly18 
      to Ala in the EGF domain of PE(delta Ia)-EGFmutants lead to a decrease in the 
      binding affinity to EGF receptor and cytotoxicity to A431 cells. In addition, 
      mutations of any of the cysteine residues of EGF in PE(delta Ia)-EGFmutants 
      resulted in the loss of their binding activity to EGF receptor and a 
      corresponding loss of their cytotoxicity. This study indicates that the 
      cytotoxicity of PE(delta Ia)-EGFmutant to EGF receptor-bearing cells may be used 
      as an indicator to screen mutations of EGF important in EGF-receptor 
      interactions.
FAU - Shiah, H S
AU  - Shiah HS
AD  - Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan, Republic of 
      China.
FAU - Chen, T Y
AU  - Chen TY
FAU - Chang, C M
AU  - Chang CM
FAU - Chow, J T
AU  - Chow JT
FAU - Kung, H J
AU  - Kung HJ
FAU - Hwang, J
AU  - Hwang J
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Bacterial Toxins)
RN  - 0 (Exotoxins)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Virulence Factors)
RN  - 62229-50-9 (Epidermal Growth Factor)
RN  - EC 2.4.2.- (ADP Ribose Transferases)
RN  - EC 2.7.10.1 (ErbB Receptors)
SB  - IM
MH  - *ADP Ribose Transferases
MH  - Amino Acid Sequence
MH  - *Bacterial Toxins
MH  - Base Sequence
MH  - Cell Line
MH  - Cell Survival/drug effects
MH  - Cloning, Molecular
MH  - Epidermal Growth Factor/genetics/*metabolism/pharmacology
MH  - ErbB Receptors/*metabolism
MH  - Escherichia coli/genetics
MH  - Exotoxins/genetics/*metabolism/pharmacology
MH  - Humans
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Plasmids
MH  - Pseudomonas aeruginosa/genetics/metabolism
MH  - Recombinant Fusion Proteins/*metabolism/pharmacology
MH  - *Virulence Factors
MH  - Pseudomonas aeruginosa Exotoxin A
EDAT- 1992/11/25 00:00
MHDA- 1992/11/25 00:01
CRDT- 1992/11/25 00:00
PHST- 1992/11/25 00:00 [pubmed]
PHST- 1992/11/25 00:01 [medline]
PHST- 1992/11/25 00:00 [entrez]
AID - S0021-9258(18)35941-6 [pii]
PST - ppublish
SO  - J Biol Chem. 1992 Nov 25;267(33):24034-40.