Abstract
The phosphatidylcholine preferring phospholipase C from Bacillus cereus (PC-PLCBc) catalyzes the hydrolysis of phospholipids in the following order of preference: phosphatidylcholine (PC) > phosphatidylethanolamine (PE) > phosphatidylserine (PS). In previous work, mutagenic, kinetic, and crystallographic experiments suggested that varying the amino acids at the 4th, 56th, and 66th positions had a significant influence upon the substrate specificity profile of PC-PLCBc. Here, we report the crystal structures of the native form of several PC-PLCBc variants that exhibited altered substrate specificities for PC, PE, and PS at maximum resolutions of 1.90-2.05 Å. Comparing the structures of these variants to the structure of the wild-type enzyme reveals only minor differences with respect to the number and location of active site water molecules and the side chain conformations of residues at the 4th and 56th positions. These results suggest that subtle changes in steric and electronic properties in the substrate binding site of PC-PLCBc are responsible for the significant changes in substrate selectivity.
| Original language | English |
|---|---|
| Pages (from-to) | 41-47 |
| Number of pages | 7 |
| Journal | Archives of Biochemistry and Biophysics |
| Volume | 460 |
| Issue number | 1 |
| DOIs | |
| State | Published - 1 Apr 2007 |
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Keywords
- Bacillus cereus
- Crystallography
- Phosphatidylcholine
- Phospholipase C
- Substrate specificity
Cite this
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Structural studies examining the substrate specificity profiles of PC-PLCBc protein variants. / Benfield, Aaron P.; Goodey, Nina; Phillips, Lauren T.; Martin, Stephen F.
In: Archives of Biochemistry and Biophysics, Vol. 460, No. 1, 01.04.2007, p. 41-47.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Structural studies examining the substrate specificity profiles of PC-PLCBc protein variants
AU - Benfield, Aaron P.
AU - Goodey, Nina
AU - Phillips, Lauren T.
AU - Martin, Stephen F.
PY - 2007/4/1
Y1 - 2007/4/1
N2 - The phosphatidylcholine preferring phospholipase C from Bacillus cereus (PC-PLCBc) catalyzes the hydrolysis of phospholipids in the following order of preference: phosphatidylcholine (PC) > phosphatidylethanolamine (PE) > phosphatidylserine (PS). In previous work, mutagenic, kinetic, and crystallographic experiments suggested that varying the amino acids at the 4th, 56th, and 66th positions had a significant influence upon the substrate specificity profile of PC-PLCBc. Here, we report the crystal structures of the native form of several PC-PLCBc variants that exhibited altered substrate specificities for PC, PE, and PS at maximum resolutions of 1.90-2.05 Å. Comparing the structures of these variants to the structure of the wild-type enzyme reveals only minor differences with respect to the number and location of active site water molecules and the side chain conformations of residues at the 4th and 56th positions. These results suggest that subtle changes in steric and electronic properties in the substrate binding site of PC-PLCBc are responsible for the significant changes in substrate selectivity.
AB - The phosphatidylcholine preferring phospholipase C from Bacillus cereus (PC-PLCBc) catalyzes the hydrolysis of phospholipids in the following order of preference: phosphatidylcholine (PC) > phosphatidylethanolamine (PE) > phosphatidylserine (PS). In previous work, mutagenic, kinetic, and crystallographic experiments suggested that varying the amino acids at the 4th, 56th, and 66th positions had a significant influence upon the substrate specificity profile of PC-PLCBc. Here, we report the crystal structures of the native form of several PC-PLCBc variants that exhibited altered substrate specificities for PC, PE, and PS at maximum resolutions of 1.90-2.05 Å. Comparing the structures of these variants to the structure of the wild-type enzyme reveals only minor differences with respect to the number and location of active site water molecules and the side chain conformations of residues at the 4th and 56th positions. These results suggest that subtle changes in steric and electronic properties in the substrate binding site of PC-PLCBc are responsible for the significant changes in substrate selectivity.
KW - Bacillus cereus
KW - Crystallography
KW - Phosphatidylcholine
KW - Phospholipase C
KW - Substrate specificity
UR - http://www.scopus.com/inward/record.url?scp=33947573496&partnerID=8YFLogxK
U2 - 10.1016/j.abb.2007.01.023
DO - 10.1016/j.abb.2007.01.023
M3 - Article
VL - 460
SP - 41
EP - 47
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
SN - 0003-9861
IS - 1
ER -