Georg o Papad Akou

7/25/2019 Georg o Papad Akou

1/5

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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2/5

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


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


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 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


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 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 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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3/5

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


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


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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4/5

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


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 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


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 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


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


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


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


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 r o t e i n

f o r

, B - l a c t a m


i n

a c l i n i c a l i s o l a t e

o f


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


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


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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.org/

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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


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


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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