georg o papad akou
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7/25/2019 Georg o Papad Akou
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22(1):172. DOI: 10.1128/AAC.22.1.172.
1982,Antimicrob. Agents Chemother.P BonnerN H Georgopapadakou, S A Smith and Dbeta-lactam antibiotics.strain resistant to specific
a Staphylococcus aureusPenicillin-binding proteins in
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V o l . 2 2 , N o .
1
N T I M I C R O B I A L
AGENTS
AND
CHEMOTHERAPY,
J u l y
1 9 8 2 ,
p .
1 7 2 - 1 7 5
0 0 6 6 - 4 8 0 4 / 8 2 / 0 7 0 1 7 2 - 0 4 0 2 . 0 0 / 0
P e n i c i l l i n - B i n d i n g P r o t e i n s
i n a
S t a p h y l o c o c c u s
a u r e u s S t r a i n
R e s i s t a n t
t o
S p e c i f i c
, - L a c t a m
A n t i b i o t i c s
NAFSIKA H .
G E O R G O P A P ADAKOU,*
S A N D R A A . SMITH,
AND
DANIEL
P .
BONNER
T h e
S q u i b b I n s t i t u t e
f o r
M e d i c a l R e s e a r c h ,
P r i n c e t o n
N ew J e r s e y
0 8 5 4 0
R e c e i v e d
2 8
D e c e m b e r
1 9 8 1 / A c c e p t e d
2 0 A p r i l 1 9 8 2
T h e
p e n i c i l l i n - b i n d i n g
p r o t e i n s
( P B P s ) o f
a
c l i n i c a l i s o l a t e o f
S t a p h y l o c o c c u s
a u r e u s
s p e c i f i c a l l y r e s i s t a n t
t o o r a l c e p h a l o s p o r i n s w e r e c o m p a r e d
w i t h t h o s e o f a
s u s c e p t i b l e
s t r a i n . I n t h e r e s i s t a n t
s t r a i n , -
PBP3 ( 7 5 , 0 0 0
m o l e c u l a r
w e i g h t ) w a s
m i s s i n g
o r h a d s u b s t a n t i a l l y ( > 1 0 0 - f o l d )
r e d u c e d
a f f i n i t y f o r
p e n i c i l l i n ;
PBP2
( 8 0 , 0 0 0
m o l e c u l a r w e i g h t ) w a s
i n c r e a s e d i n a m o u n t a n d c o n t a i n e d a s a t e l l i t e
b a n d ,
P B P 2 ; P B P s
1
a n d
4 w e r e
u n c h a n g e d .
O r a l c e p h a l o s p o r i n s
b o u n d
p o o r l y
t o
PBP2
i n
b o t h
s u s c e p t i b l e
a n d
r e s i s t a n t
s t r a i n s ,
b u t o n l y
i n
t h e
l a t t e r
d i d
b i n d i n g
c o r r e l a t e
w i t h a n t i b i o t i c a c t i v i t y . T h e
r e s u l t s a r e
c o n s i s t e n t
w i t h t h e s u g g e s t i o n t h a t
PBP2
i s
e s s e n t i a l i n
S .
a u r e u s .
PBP2 m i g h t
i n
a d d i t i o n
c o m p e n s a t e f o r P B P 3 w h e n
t h e
l a t t e r i s
m i s s i n g .
I n
t h e s u s c e p t i b l e s t r a i n
t h e l a c k
o f c o r r e l a t i o n
b e t w e e n b i n d i n g
t o P B P 2
a n d
P - l a c t a m
a n t i b i o t i c a c t i v i t y
i s d u e
t o
t h e
v e r y
h i g h
a f f i n i t y o f t h e a l s o
e s s e n t i a l P B P 3
f o r
P - l a c t a m
a n t i b i o t i c s .
C e p h r a d i n e - r e s i s t a n t
s t r a i n s
o f
S t a p h y l o c o c -
c u s a u r e u s
h a v e
b e e n
r e c e n t l y
d e s c r i b e d
8 ) .
R e s i s t a n c e
w a s
r e p o r t e d
t o b e
i n t r i n s i c r a t h e r
t h a n d u e
t o
t h e
p r e s e n c e
o f
1 - l a c t a m a s e s
a n d ,
i n
c o n t r a s t
t o
m e t h i c i l l i n
r e s i s t a n c e ,
t o
b e r e s t r i c t -
e d
t o
o r a l
c e p h a l o s p o r i n s .
I n t r i n s i c
b a c t e r i a l
r e s i s t a n c e t o
1 3 - l a c t a m
a n t i -
b i o t i c s
h a s b e e n
s h o w n t o
b e d u e
t o
c h a n g e s
i n
t h e
a m o u n t s
( 1 )
o r a f f i n i t i e s
f o r
P - l a c t a m
a n t i b i -
o t i c s
( 3 , 6 )
o f
e s s e n t i a l
p e n i c i l l i n - b i n d i n g
p r o -
t e i n s
( P B P s ) .
P B P s a r e
c y t o p l a s m i c
m e m b r a n e
p r o t e i n s ( u s u a l l y
f i v e t o
e i g h t ,
o f
m o l e c u l a r
w e i g h t s 4 0 , 0 0 0
[ 4 0 K ]
t o 1 0 0 K )
w h i c h s p e c i f i c a l l y
b i n d
p e n i c i l l i n
a n d o t h e r
, - l a c t a m
a n t i b i o t i c s
a n d
a r e i n v o l v e d
i n
b a c t e r i a l
c e l l
w a l l
b i o s y n t h e -
s i s . E s s e n t i a l P B P s
a r e
t h o s e
( u s u a l l y
o f
m o l e c u -
l a r
w e i g h t s
7 0
t o
1 0 0 K )
w h o s e
b i n d i n g
t o
P -
l a c t a m
a n t i b i o t i c s
c o r r e l a t e s
w i t h a n t i b i o t i c
a c t i v i t y 2 , 1 3 , 1 6 ) .
I n
t h e c a s e
o f
E s c h e r i c h i a
c o l i a n d
r e l a t e d
b a c t e r i a ,
e s s e n t i a l
PBPs a r e
a s s o c i a t e d
w i t h
i m p o r t a n t
p h y s i o l o g i c a l
f u n c -
t i o n s s u c h
a s
e l o n g a t i o n
a n d
s e p t a t i o n ( 1 5 ) .
I n
S .
a u r e u s f o u r P B P s
h a v e
b e e n
d e t e c t e d ,
o f
w h i c h P B P 3 ( 3 )
a n d
PBP2
( 6 )
a r e
e s s e n t i a l .
U n f o r t u n a t e l y ,
w i t h
t h e
e x c e p t i o n
o f
t h e
4 1 K
p r o t e i n
( D D - c a r b o x y p e p t i d a s e
[ 7 ] ) ,
n o n e
h a s d e -
m o n s t r a b l e e n z y m a t i c
a c t i v i t y .
I n t h e
p r e s e n t
s t u d y
t h e PBPs o f
a
c e p h r a d i n e - r e s i s t a n t
c l i n i c a l
i s o l a t e
o f
S .
a u r e u s
w e r e
c o m p a r e d
w i t h
t h o s e
o f
a
c e p h r a d i n e - s u s c e p t i b l e
s t r a i n .
An
e x p l a n a t i o n
f o r
t h i s
t y p e
o f
i n t r i n s i c
r e s i s t a n c e , a p p a r e n t l y
c o n f i n e d t o a
r a t h e r s m a l l
n u m b e r
o f
P - l a c t a m
a n t i b i o t i c s ,
i s
g i v e n ,
a n d r e s i s t a n c e t o c o m -
p o u n d s
o t h e r
t h a n
o r a l
c e p h a l o s p o r i n s
i s
p r e -
d i c t e d
a n d
c o n f i r m e d .
A
c e p h r a d i n e - r e s i s t a n t
c l i n i c a l
i s o l a t e o f
S .
a u r e u s ( S q u i b b C u l t u r e
[ S C ]
1 2 , 7 0 0 )
w a s k i n d l y
p r o v i d e d b y
R .
W.
L a c e y ,
Q u e e n E l i z a b e t h
H o s p i t a l ,
N o r f o l k ,
E n g l a n d .
S i n c e
a c e p h r a d i n e -
s u s c e p t i b l e
s t r a i n i s o g e n i c
t o
S C 1 2 , 7 0 0
i s
n o t
a v a i l a b l e ,
S .
a u r e u s SC
2 3 9 9 ,
a
s t r a i n w i t h
t y p i c a l , - l a c t a m
s u s c e p t i b i l i t y
a n d P B P
p r o f i l e
( 3 ) ,
w a s u s e d a s t h e
c o n t r o l
s t r a i n .
B o t h o r g a
n i s m s
w e r e g r o w n
i n
b r a i n
h e a r t
i n f u s i o n m e d i -
um
( D i f c o L a b o r a t o r i e s ,
D e t r o i t ,
M i c h . )
a t
3 7 C
o n a
G y r o t o r y
s h a k e r
( 2 8 0 r p m ;
N ew B r u n s w i c k
S c i e n t i f i c
C o . ,
N ew B r u n s w i c k ,
N . J . )
f o r 1 6
h
a n d
s u b s e q u e n t l y
( 1 0
i n o c u l u m ) f o r
4
h .
C e l l s
w e r e h a r v e s t e d b y
c e n t r i f u g a t i o n
a n d
s o n i c a t e d
w i t h c o o l i n g ( i c e - s a l t
b a t h ) f o r 9 m i n ,
a n d
mem-
b r a n e s
w e r e s o l u b i l i z e d
i n
2 T r i t o n
X - 1 0 0
a s
p r e v i o u s l y
d e s c r i b e d
3 ) .
A l t e r n a t i v e l y ,
h a r v e s t -
e d c e l l s w e r e i n c u b a t e d
w i t h
l y s o s t a p h i n
( 5 0
, u g /
m l )
a n d
l y s o z y m e ( 1 0 0 , u g / m l )
f o r 3 0
m i n
a t
3 7 C
i n 5 0 mM
p o t a s s i u m p h o s p h a t e ( p H 7 . 5 ) - 4
M
N a C l ,
t h e n
l y s e d
i n 1 0 mM
p o t a s s i u m p h o s p h a t e
( p H 7 . 5 ) - 2
mM
M g C l 2
c o n t a i n i n g
2
, u g
o f
DNase
p e r
m l
( 1 0 ) .
M e m b r a n e s w e r e i n c u b a t e d
( 1 0 0 , u g
o f
p r o t e i n ,
d e t e r m i n e d
b y
t h e
m e t h o d
o f
L o w r y
e t
a l .
[ 1 2 ] )
w i t h t h e
a p p r o p r i a t e
a m o u n t
o f
I -
l a c t a m
a n t i b i o t i c
( f r e s h l y
p r e p a r e d
a q u e o u s
s o -
l u t i o n ;
f i n a l
c o n c e n t r a t i o n s ,
0 ,
0 . 1 , 0 . 5 , 2 . 0 , 1 0 ,
3 0 ,
a n d 1 0 0
, L g / m l )
a t
3 0 C
f o r
1 0
m i n
i n
a
t o t a l
v o l u m e
o f
5 0
R I .
[ 8 - 1 4 C ] p e n i c i l l i n
G
( 1 0
n m o l ;
A m e r s h a m C o r p ; s p e c i f i c a c t i v i t y ,
5 4
, u C i / , m o l )
w a s
a d d e d ,
a n d t h e i n c u b a t i o n
was
c o n t i n u e d f o r
a n o t h e r
1 0 m i n . P r o t e i n
was
p r e c i p i t a t e d
w i t h
4
v o l u m e s o f
c o l d a c e t o n e
a n d
s u b j e c t e d
t o s o d i -
um
d o d e c y l s u l f a t e - p o l y a c r y l a m i d e
g e l
e l e c t r o -
p h o r e s i s
( 9 ;
r u n n i n g
g e l ,
1 0
a c r y l a m i d e - 0 . 2 7
1 7 2
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N O T E S 1 7 3
1 . 0 - R 1 - 1
~
R
-
---I
K)
0 . . .. . S
-
F I G . 1 .
B i n d i n g
o f
[ 4 C ] b e n z y l p e n i c i l l i n
t o
m e m b r a n e s
o f
c e p h r a d i n e - s u s c e p t i b l e
( B )
a n d
- r e s i s t a n t
( A )
S .
aureus
s t r a i n s . A n t i b i o t i c c o n c e n t r a t i o n s ( m i c r o g r a m s per m i l l i l i t e r )
are i n d i c a t e d .
b i s - a c r y l a m i d e )
f o l l o w e d b y f l u o r o g r a p h y
( 1 1 ) .
P r o t e i n - b o u n d
[ 1 4 C ] p e n i c i l l i n
G was
d e t e r m i n e d
b y
v i s u a l
e x a m i n a t i o n
o f
t h e
X - r a y
f i l m .
M e m b r a n e s
p r e p a r e d f r o m
s o n i c a t e d
c e l l s
or
l y s e d s p h e r o p l a s t s
gave
i d e n t i c a l
r e s u l t s . S o l u b i -
l i z a t i o n
o f m e m b r a n e s b y T r i t o n
X - 1 0 0
d i d n o t
a f f e c t
PBPs
e i t h e r ,
a n d
s o l u b i l i z e d
m e m b r a n e s
were
r o u t i n e l y
u s e d f o r
t h e
s a k e
o f
c o n v e n i e n c e .
Minimum
i n h i b i t o r y
c o n c e n t r a t i o n s
were d e -
t e r m i n e d
b y
t h e t u b e
d i l u t i o n
m e t h o d
( t w o f o l d
s e r i a l
d i l u t i o n s
f r o m
1 0 0 t o
0 . 0 5 , u g / m l )
i n A n t i b i -
o t i c
A s s a y
B r o t h
B BL
M i c r o b i o l o g y S y s t e m s ,
C o c k e y s v i l l e ,
M d . )
a t
a n
i n o c u l u m
o f 1 0 6
c o l o -
n y - f o r m i n g
u n i t s
per
m l .
T h e
P B P
p a t t e r n
o f t h e
c e p h r a d i n e - r e s i s t a n t
s t r a i n ,
SC
1 2 , 7 0 0 ,
was
s u b s t a n t i a l l y
d i f f e r e n t
f r o m t h a t
o f
t h e s u s c e p t i b l e
s t r a i n ,
SC 2 3 9 9 ( F i g .
1 ) .
PBP3 wa s a b s e n t
( o r h a d s u b s t a n t i a l l y r e -
d u c e d a f f i n i t y
f o r
p e n i c i l l i n
G ) ,
a n d
P B P 3 ,
a
p r o t e i n
o f m o l e c u l a r
w e i g h t
l o w e r
t h a n t h a t
o f
P B P 3 , a p p e a r e d. U n f o r t u n a t e l y , P B P 3 was n o t
c o n s i s t e n t l y p r e s e n t
( t h e r m o l a b i l e ? )
f o r
i t s
1
l a c t a m p r o f i l e
t o
b e
d e t e r m i n e d . P B P 2
was
i n -
c r e a s e d
i n
amount
a n d
h a d a
s a t e l l i t e
b a n d ,
P B P 2 ,
o f
s l i g h t l y
l o w e r
m o l e c u l a r
w e i g h t .
P B P1
a n d
4
( D D - c a r b o x y p e p t i d a s e )
were
u n c h a n g e d ,
a l t h o u g h
PBP1
was s o m e w h a t
more
t h e r m o l a -
b i l e
i n t h e
r e s i s t a n t s t r a i n .
T h e
P B P
b i n d i n g p r o f i l e s
o f
s t r u c t u r a l l y
d i -
verse
1 - l a c t a m
a n t i b i o t i c s
were i d e n t i c a l f o r
b o t h
s t r a i n s
( T a b l e s
1
a n d
2 ) .
G e n e r a l l y ,
com-
p o u n d s
w i t h
g o o d
a n t i b a c t e r i a l
a c t i v i t y
b o u n d
t o
P B P 2 ,
o r a l c e p h a l o s p o r i n s a n d m e c i l l i n a m
b e i n g
e x c e p t i o n s .
I n
SC
1 2 , 7 0 0 ,
b u t
n o t i n SC 2 3 9 9 ,
poor
P BP2
b i n d i n g
o f
c e p h r a d i n e
( a n d
o t h e r
o r a l
c e p h a l o s p o r i n s )
was a s s o c i a t e d
w i t h
r e s i s t a n c e .
M e c i l l i n a m ,
a l t h o u g h s t r u c t u r a l l y
d i f f e r e n t
f r o m
o r a l
c e p h a l o s p o r i n s ,
a l s o
b o u n d
p o o r l y
t o
P B P 2 .
A c c o r d i n g l y ,
SC
1 2 , 7 0 0 was
a l s o
r e s i s t a n t t o
m e c i l l i n a m .
, B - L a c t a m a n t i b i o t i c s a c t i v e
a g a i n s t
S . aureus
a l m o s t
a l w a y s
b i n d t o
P B P 3 ,
a n d u s u a l l y t o
T A B L E 1 .
B i n d i n g o f l a c t a m a n t i b i o t i c s t o
P B P s
o f S . a u r e u s SC 2 3 9 9
C o n c n
( , u g / m l )
t o
c o m p l e t e l y
i n h i b i t p e n i c i l l i n G b i n d i n g
M I C a
P B P 1
( 8 7 K )
PBP2
( 8 0 K ) P B P 3 ( 7 5 K ) P B P 4
( 4 1 K ) , u g / m l )
A m p i c i l l i n
0 . 1 0 . 1
0 . 1 > 1 0 0 < 0 . 0 5
C a r b e n i c i l l i n 0 . 5
0 . 5
1 0
> 1 0 0 0 . 8
C e f a c l o r
0 . 1
> 1 0 0
0 . 1
> 1 0 0 1 . 6
C e f o t a x i m e
0 . 5
0 . 5
1 0
> 1 0 0 1 . 6
C e f o x i t i n
1 0
2 . 0
1 0 0 . 5
0 . 8
C e p h a d r o x i l
0 . 1
> 1 0 0 0 . 1 > 1 0 0 0 . 8
C e p h a l e x i n
0 . 5
> 1 0 0
0 . 1
> 1 0 0 0 . 8
C e p h a l o r i d i n e
0 . 1
0 . 1
0 . 1
1 0 0
< 0 . 0 5
C e p h a l o t h i n
0 . 5
0 . 5 0 . 5 1 0 0 0 . 1
C e p h r a d i n e
0 . 5
> 1 0 0
0 . 1
> 1 0 0 0 . 8
M e c i l l i n a m
1 0 0 > 1 0 0
2 . 0 > 1 0 0 1 2 . 5
M e t h i c i l l i n 0 . 5
1 0
0 . 1
1 0 1 . 6
P e n i c i l l i n G 0 . 1
0 . 1 0 . 1 . 1 0 0 < 0 . 0 5
a
M I C ,
Minimum
i n h i b i t o r y
c o n c e n t r a t i o n .
V O L .
2 2 ,
1 9 8 2
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A N T I M I C R O B .
A G E N T S C H E M O T H E R .
T A B L E 2 . B i n d i n g o f 1 - l a c t a m
a n t i b i o t i c s
t o
P B P s
o f
S . a u r e u s
SC 1 2 , 7 0 0
Compound
C o n c n
I L g / m l )
t o
c o m p l e t e l y
i n h i b i t
p e n i c i l l i n
G
b i n d i n g
P B P 1 ( 8 7 K ) P B P 2
( 8 0 K )
P B P 2
( 7 7 K )
P B P 4
( 4 1 K )
I L g / m l )
A m p i c i l l i n
0 . 1
0 . 1 0 . 1
> 1 0 0
< 0 . 0 1
C a r b e n i c i l l i n
0 . 5
0 . 5
0 . 5
> 1 0 0 1 . 6
C e f a c l o r
0 . 1
> 1 0 0
0 . 1
> 1 0 0
1 2 . 5
C e f o t a x i m e
0 . 5
0 . 5
1 0 . 0
- 1 0 0
1 . 6
C e f o x i t i n 1 0
2 . 0
1 0 . 0
0 . 5
0 . 8
C e p h a d r o x i l
0 . 1 > 1 0 0
2 . 0 >100
1 2 . 5
C e p h a l e x i n
0 . 5
> 1 0 0 0 . 1
> 1 0 0 1 2 . 5
C e p h a l o r i d i n e
0 . 1
0 . 1 0 . 5
1 0 0
< 0 . 0 5
C e p h a l o t h i n
0 . 5
0 . 5
0 . 1 1 0 0 0 . 8
C e p h r a d i n e
0 . 5 > 1 0 0
1 0 . 0
> 1 0 0
2 5
M e c i l l i n a m 1 0 0
> 1 0 0
0 . 5
> 1 0 0 7 5
M e t h i c i l l i n
0 . 5 1 0
0 . 1
> 1 0 0
3 . 1
P e n i c i l l i n G
0 . 1
0 . 1
b
1 0 0
< 0 . 0 5
a
M I C ,
M i n i m u m
i n h i b i t o r y c o n c e n t r a t i o n .
b
No d a t a .
P B P 2 . I t i s t h e r e f o r e
d i f f i c u l t
t o
d e t e r m i n e
w h e t h e r P B P 2 i s
e s s e n t i a l
i n t h e w i l d - t y p e
s t r a i n .
W i t h
t h e e m e r g e n c e
o f m o n o b a c t a m s
( 1 7 ) , c o m p o u n d s
h a v e
b e e n f o u n d w h i c h
b i n d
p r e f e r e n t i a l l y
t o
PBP2 a n d
p o s s e s s
a n t i - s t a p h y -
l o c o c c a l a c t i v i t y ( G e o r g o p a p a d a k o u
a n d
S m i t h ,
u n p u b l i s h e d d a t a ) .
On
t h e o t h e r
h a n d ,
c e p h r a -
d i n e ,
w h i c h
b i n d s
t o
P B P 3
b u t
n o t
t o
P B P 2 ,
i s
a l s o
a c t i v e a g a i n s t S .
a u r e u s . T h e r e f o r e ,
a s
s u g g e s t e d
3 ,
6 ) ,
P B P s 2 a n d 3
a r e
e s s e n t i a l
a n d
d o
n o t
n o r m a l l y
i n t e r c o m p e n s a t e .
I n
t h e
c e p h r a -
d i n e - r e s i s t a n t
s t r a i n
SC 1 2 , 7 0 0 ,
P B P 3
h a s
r e -
d u c e d
a f f i n i t y
f o r
b e n z y l p e n i c i l l i n
( n o b i n d i n g
a t
1 , 0 0 0
, u g l m l )
o r i s a l t o g e t h e r
m i s s i n g .
I n
e i t h e r
c a s e , s u s c e p t i b i l i t y
o r
r e s i s t a n c e
t o
P - l a c t a m
a n t i b i o t i c s
a p p e a r s
t o b e
s o l e l y
d e t e r m i n e d
b y
b i n d i n g
t o P B P 2 .
T h e o b s e r v e d
i n c r e a s e
i n t h e
a m o u n t
o f
PBP2
a n d
a p p e a r a n c e
o f P B P s
2 a n d
3
ma y
b e
c o m p e n s a t o r y
f o r P B P 3 .
S u c h c o m -
p e n s a t o r y
c h a n g e s
h a v e b e e n
o b s e r v e d
w i t h
P B P s o f B a c i l l u s
m e g a t e r i u m
a n d
S t r e p t o c o c c u s
p n e u m o n i a e
s t r a i n s
r e s i s t a n t
t o
p - l a c t a m
a n t i b i -
o t i c s
4 , 5 ,
1 4 ) .
P B P s
2 a n d 3 o f S .
a u r e u s
ma y
b e
b o t h
p e p t i d o g l y c a n
t r a n s p e p t i d a s e s
t h a t h a v e
d i f f e r e n t
p h y s i o l o g i c a l
f u n c t i o n s ,
b u t
a r e
s u f f i -
c i e n t l y
s i m i l a r
t o
i n t e r c o m p e n s a t e
u n d e r
a n t i b i -
o t i c
p r e s s u r e .
I n
c o n c l u s i o n ,
a
c e p h r a d i n e - r e s i s t a n t
c l i n i c a l
i s o l a t e
o f S .
a u r e u s
l a c k s
PBP3
a n d
h a s
( c o m -
p e n s a t o r y ? )
c h a n g e s
i n o t h e r
P B P s . R e s i s t a n c e
i s
c o n f i n e d
t o
a s m a l l
n u m b e r
o f
c o m p o u n d s ,
m o s t
s t r u c t u r a l l y r e l a t e d ,
a l l
s h o w i n g p o o r
b i n d -
i n g
t o
P B P 2 .
A n t i b i o t i c a c t i v i t y
i s
a s s o c i a t e d
w i t h
b i n d i n g
t o
P B P 2 .
F u r t h e r
s t u d i e s a r e
n e e d -
e d
t o e l u c i d a t e
t h e
p h y s i o l o g i c a l
f u n c t i o n s
a n d
e n z y m a t i c
a c t i v i t i e s
o f
P B P s
2 a n d
3 .
E q u a l l y
i m p o r t a n t ,
a d d i t i o n a l
c e p h r a d i n e - r e s i s t a n t
s t r a i n s m u s t b e
e x a m i n e d
t o e s t a b l i s h
t h e
p r e v a -
l e n c e
o f t h i s
p a t t e r n
o f
r e s i s t a n c e .
We t h a n k B e a t r i c e
F . M i n a s s i a n f o r
s u p p l y i n g
t h e m i n i m u m
i n h i b i t o r y c o n c e n t r a t i o n
d a t a ,
a n d
R i c h a r d B . S y k e s f o r
h i s
e n c o u r a g e m e n t
a n d f o r
c r i t i c a l l y r e a d i n g t h e
m a n u s c r i p t .
LITERATURE
CITED
1 .
B r o w n , D .
F . J . , a n d P .
R e y n o l d s . 1 9 8 0 . I n t r i n s i c
r e s i s t -
a n c e t o
, - l a c t a m
a n t i b i o t i c s
i n S t a p h y l o c o c c u s
a u r e u s .
FEBS
L e t t .
1 2 2 : 2 7 5 - 2 8 0 .
2 .
D o u g h e r t y ,
T .
J . ,
A .
E . K o l l e r , a n d A .
T o m s z .
1 9 8 0 .
P e n i c i l l i n - b i n d i n g p r o t e i n s
o f
p e n i c i l l i n - s u s c e p t i b l e
a n d
i n t r i n s i c a l l y
r e s i s t a n t N e i s s e r i a g o n o r r h o e a e .
A n t i m i c r o b .
A g e n t s C h e m o t h e r .
1 8 : 7 3 0 - 7 3 5 .
3 .
G e o r g o p a p a d a k o u ,
N .
H . ,
a n d
F . Y . L i u . 1 9 8 0 .
B i n d i n g o f
P - l a c t a m
a n t i b i o t i c s t o p e n i c i l l i n - b i n d i n g
p r o t e i n s o f
S t a p h y l o c o c c u s
a u r e u s a n d
S t r e p t o c o c c u s
f a e c a l i s :
r e l a -
t i o n t o a n t i b a c t e r i a l
a c t i v i t y . A n t i m i c r o b .
A g e n t s
C h e m o -
t h e r .
1 8 : 8 3 4 - 8 3 6 .
4 .
G i e s ,
A .
F . ,
a n d
P .
E .
R e y n o l d s .
1 9 7 9 .
B a c i l l u s
m e g a t e r -
i u m
r e s i s t a n c e t o
c l o x a c i l l i n
a c c o m p a n i e d b y
a c o m p e n s a -
t o r y c l e a v a g e i n p e n i c i l l i n
b i n d i n g
p r o t e i n s .
N a t u r e
( L o n -
d o n )
2 8 0 : 1 6 7 - 1 6 8 .
5 .
H a k e n b e c k ,
R . ,
M.
T a r p a y , a n d
A .
T o m a s z .
1 9 8 0 .
M u l t i p l e
c h a n g e s
o f
p e n i c i l l i n - b i n d i n g p r o t e i n s
i n
p e n i c i l l i n - r e s i s t -
a n t
c l i n i c a l
i s o l a t e s o f S t r e p t o c o c c u s p n e u m o n i a e .
A n t i -
m i c r o b .
A g e n t s
C h e m o t h e r .
1 7 : 3 6 4 - 3 7 1 .
6 .
H a y e s ,
M.
V . ,
N . A .
C .
C u r t i s ,
A . W.
W y k e ,
a n d
J .
B .
W a r d . 1 9 8 1 .
D e c r e a s e d a f f i n i t y
o f a
p e n i c i l l i n - b i n d i n g
p r o t e i n
f o r
, B - l a c t a m
a n t i b i o t i c s
i n
a c l i n i c a l i s o l a t e
o f
S t a p h y l o c o c c u s
a u r e u s
r e s i s t a n t t o
m e t h i c i l l i n . F E M S
M i c r o b i o l .
L e t t .
1 0 : 1 1 9 - 1 2 2 .
7 .
K o z a r c h ,
J .
W . ,
a n d
J .
L .
S t r o n i n g e r .
1 9 7 8 .
m e m b r a n e
e n z y m e
f r o m
S t a p h y l o c o c c u s
a u r e u s
w h i c h
c a t a l y z e s
t r a n s p e p t i d a s e ,
c a r b o x y p e p t i d a s e
a n d
p e n i c i l l i n a s e
a c t i v i -
t i e s . J . B i o l .
C h e m . 2 5 3 : 1 2 7 2 - 1 2 7 8 .
8 .
L a c e y ,
R .
W . ,
a n d
V .
L o r d .
1 9 8 1 .
N ew
t y p e
o f 1 - l a c t a m
r e s i s t a n c e
i n
S t a p h y l o c o c c u s
a u r e u s .
L a n c e t
1 : 1 0 4 9 - 1 0 5 0 .
9 .
L a e m m l l ,
U .
K .
1 9 7 0 .
C l e a v a g e
o f
s t r u c t u r a l
p r o t e i n s
d u r i n g
t h e
a s s e m b l y
o f
t h e
h e a d o f
b a c t e r i o p h a g e
T 4 .
N a t u r e
( L o n d o n )
2 2 7 : 6 8 0 - 6 8 5 .
1 0 .
L a s c e l l e s ,
J .
1 9 7 9 . H e m e - d e f i c i e n t
m u t a n t s
o f
S t a p h y l o -
c o c c u s
a u r e u s .
M e t h o d s
E n z y m o l .
5 6 : 1 7 2 - 1 8 0 .
1 1 .
L a s k e y ,
R .
A . ,
a n d
A . D . M i l l s . 1 9 7 5 .
Q u a n t i t a t i v e
f i l m
d e t e c t i o n o f
3 H
a n d
4 C i n
p o l y a c r y l a m i d e
g e l s
b y f l u o r o g -
r a p h y .
E u r .
J .
B i o c h e m .
5 6 : 3 3 5 - 3 4 1 .
1 7 4
N O T E S
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V O L . 2 2 ,
1 9 8 2
NOTES
1 7 5
1 2 . L o w r y , 0 . H . ,
N . J .
R o s e b r o u g h , A .
L .
F a r r , a n d R . J .
R a n d a l l . 1 9 5 1 . P r o t e i n
m e a s u r e m e n t w i t h t h e F o l i n
p h e n o l
r e a g e n t . J . B i o l .
C h e m .
1 9 3 : 2 6 5 - 2 7 5 .
1 3 .
O g a w a r a , H . , a n d S .
H o r i l c a w a . 1 9 8 0 .
P e n i c i l l i n - b i n d i n g
p r o t e i n s o f
S t r e p t o m y c e s
c a c a o i ,
S t r e p t o m y c e s o l i v a c e u s
a n d
S t r e p t o m y c e s
c l a v u l i g e r u s .
A n t i m i c r o b .
A g e n t s
C h e m o t h e r .
1 7 : 1 - 7 .
1 4 . R e y n o l d s , P .
E . , S .
T .
S h e p h e r d ,
a n d H .
A . C h a s e . 1 9 7 8 .
I d e n t i f i c a t i o n o f t h e
b i n d i n g p r o t e i n
w h i c h m a y
b e t h e
t a r g e t
o f
p e n i c i l l i n a c t i o n i n
B a c i l l u s m e g a t e r i u m .
N a t u r e
( L o n d o n ) 2 7 1 : 5 6 8 - 5 7 0 .
1 5 .
S p r a t t ,
B .
G . 1 9 7 5 .
D i s t i n c t p e n i c i l l i n - b i n d i n g
p r o t e i n s
i n v o l v e d i n t h e
d i v i s i o n , e l o n g a t i o n
a n d s h a p e o f
E s c h e -
r i c h i a c o l i
K 1 2 .
P r o c . N a t l . A c a d .
S c i .
U . S . A . 7 2 : 2 9 9 9 -
3 0 0 3 .
1 6 . S p r a t t ,
B .
G . 1 9 7 8 .
E s c h e r i c h i a
c o l i
r e s i s t a n c e
t o
P - l a c t a m
a n t i b i o t i c s
t h r o u g h
a d e c r e a s e i n
t h e
a f f i n i t y
o f a
t a r g e t f o r
l e t h a l i t y .
N a t u r e
( L o n d o n )
2 7 4 : 7 1 3 - 7 1 5 .
1 7 .
S y k e s , R .
B . , C .
M.
C i m a r u s t ,
D .
P . B o n n e r ,
K .
B u s h ,
D . M . F l o y d ,
N . H .
G e o r g o p a p a d a k o u , W. H . K o s t e r ,
W.
C .
L i n ,
W. L .
P a r k e r ,
P .
A . P r i n dp e ,
M . L .
R a t h n u m ,
W.
A .
S l u s a r c h y k ,
W.
H .
T r e j o ,
a n d J .
S .
W e D s . 1 9 8 1 .
M o n o c y c l i c
P - l a c t a m
a n t i b i o t i c s
p r o d u c e d b y b a c t e r i a .
N a t u r e
( L o n d o n ) 2 9 1 : 4 8 9 - 4 9 1 .
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