An ESR study of pseudomonas copper-protein

Vol. 10, No. 1, 1963

AN

BIOCHEMICAL

ESR

STUDY

OF

AND BIOPHYSICAL

PSEUDOMONAS

H.

Department

Received

of

November

of

tionally

low

in

and

lactase

not

and

one

of

and

suggested

unique

manner (1)

unpaired

two

and

spin

resonance

one Blue

6) and

This

Portland

in

close

that

study

pairs

we

electron

Cu

(II)

between

two

copper

that

it

was

the

,

coordiof the

Cu

(I),

but

nuclei,

spin

resonance

“similar structure

in

and

-1

cm

is

between

proximity

of

copper

atom

microorganisms

was supported United States

been

investigated

may

of the

be used

resonance

per

atoms

has

have

spin

copper-proteins

study and the indebted.

copper

electron

excep-

0.008

delocalization

hyperfine

copper-oxidases

copper

other

and

the

resonance

have

of

the

of nuclear

spin to

-1

cm

degree

examined found

them

enzymes

equally

They

are

these

electron

found

= 0.009

interaction

lack

the

a high

shared

spectroscopy the

--only

School,

to

spec-

that

of

indicates

enzyme

and

comprise

the

lactase that

undergo

at

ex-

(3).

present

determining

,+

ions

of important In the

in

associates

hypothesis

4).

(5,

his

A

involving

perhaps

The

interaction”

centers

that

oxidase.

copper

The

Medical

2) and

(1, constants,

electron

cytochrome

change

have

or

ceruloplasmin.

least

Oregon

determined

ceruloplasmin

It was

Beinert trum

of

have

splitting

electron

as

Vanngard

hyperfine

some

unpaired

Mason

28, 1962

respectively. nated

COPPER-PROTEIN*

University

Biochemistry,

Malmstrom spectra

S.

RESEARCH COMMUNICATIONS

to

considerable

interest

possibility

detect

spectra

prosthetic

such of two

that

(cf.

el’ectron

structures

by

copper-proteins

which

molecule. have

been (7).

isolated They

from have

by grants from the Public Health Service,

11

various

pseudomonads

unusually

intense

absorp-

American to whom

Cancer we are

Society greatly

Vol. 10, No. 1, 1963

tion

in

atom

the of

region

copper

of the

blue

tained

0.76

form ESR

analysis

0.84

atom

it

by

of

a Varian

trated splitting

chemical

Dr.

per

molecule,

analysis,

and as

molecule;

chemical

second

Ambler

analysis),

0.65

one

0.67

atom

a standard. 0.76

0.70

Pseudomonas

which

atom

was by

His

second

sample

per

(10)

and

by

conthe

cupric

quantitative contained was

ESR

provided

denitrificans

in

molecule

quantitative

generously

samples

of which

molecule

by

one two

per

atom

atom

and

provided

aeruginosa, of

was

16,000,

generously

0 f this,

copper-protein

from

P.

of about

Pseudomonas

cupric-EDTA per

weights

RESEARCH COMMUNICATIONS

signals

obtained

V-4500

amplitudes by

Fig. constant

from

spectrometer between 1.

A

with 8 and

They

these

had

= 0.006

32

gmax cm

two

100

kc

gauss,

field

modulation

were

, the

almost

H. in

lowest

yet

at

-165”,

and

field

identical

g-parallel

= 2.055, -1

copper-proteins

Iwasaki; the

crys-

and

using modulaare

illus-

= 2. 157,

and

a hyperfine

for

any

copper

observed

protein.

19max

Figure 1. Electron spin protein from Pseudomonas T = -165”.

resonance aeruginosa

signal in

12

(derivative) of 0.05 M ammonium

the

spectro-

Dr.

examined

in

form. The

tion

molecular

mu,

copper

copper

isolated

AND BIOPHYSICAL

from

of

(by

The

talline

610

molecule.

using

form

was

per

atom 9)

scopy.

of

copper-protein

(8,

cupric

BIOCHEMICAL

1 mM copperacetate,

pH

6.5,

Vol. 10, No. 1, 1963

BIOCHEMICAL

These

results

explanation

for

sought

small

and

spectrum, in

that,

for

hyperfine

for

due

in

the

to

in

involves

Dr.

N.

one

Horie

RESEARCH COMMUNICATIONS

copper-proteins

of absorption which

are

some

splittings

exhaltation

a configuration

Thanks with

mean

AND BIOPHYSICAL

and

at

g-parallel the

Mr.

region

610

copper

mu

atom

Paul

any

region,

rate,

the

of the

ESR

must

be

only.

McMahill

for

assistance

analyses.

References

1.

Malmstrom,

2.

Broman, Biol.,

3.

Beinert, J.

4.

Mason,

5. 6.

B., L., -5, H.,

D.

Nature,

Horio,

T.,

J.

Biochem.,

Coval, Acta.

M.

L., -51,

Stark,

G.,

9.

Griffiths, (1961). Suzuki,

246 I.

and D.

H.,

and

Dawson, E.,

and

and

Iwasaki,

79

and

D.

195

Kamen,

2,

118

Vanngard.

C.,

Anal.

J.

C.,

Biochem.,

13

D.,

Proc.

Chem., D.

(1958);45,

M.

H.,

Wharton,

Biol.,

(1960).

J. Mol.

T.,

and

Sands,

-

2.67

R.

H.

(1956).

Wilkinson,

H.,

R.,

45, -

and

C.,

Mol.

Wharton, (1962).

Tokyo, T.,

J.

Aasa,

E., 2337

177,

Horio, (1961).

W.,

T., B.,

237,

S.,

8.

10.

Griffiths, Chem.,

H.

(1962).

Vanngard,

Malmstrom, 301 (1962).

Biol.

Sutherland, 43P

7.

and

Biochem.

Biochem.

-30, J.

Biol.

Tokyo,

191

(1958). Biophys.

Sot.,

84, -

(1958).

Chem.,

236,

52, -_

193

1857

(1962).

_