Different rates of synthesis of whey protein and casein by alleles of the β-lactoglobulin and αs1-casein locus in cattle

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Different rates of synthesis of whey protein and casein

by alleles of the β-lactoglobulin and αs1-casein locus in

cattle

R. Graml, G. Weiss, J. Buchberger, F. Pirchner

To cite this version:

Note

Different

rates

of

_synthesis

of

_whey

_protein

and

casein

_by

alleles

of

the

_{β-lactoglobulin}

and

_α

_s1

_-casein

locus

in

cattle

R. Graml

G.

Weiss

2

J.

_Buchberger

₂

F. Pirchner

1

1

Lehrstuhl für

Tierzucht der Technischen Universität _München, D-8050

Freising-Weihen-stephan;

2

Institut _für Chemie und _Physik der Süddeutschen Versuchs- und _{Forschungsanstalt für}

Milchwirtschaft der Technischen Universität München, D-8050 _{Freising-Weihenstephan,} FRG

(received

22 March _{1989; accepted} 28

_{August 1989)}

Summary - Quantities of cx"1-caseins and _{#-lactoglobulins} were determined in milk of 2059 Fleckvieh cows and 1809 Brnunvieh cows in _Bavaria; 6353 milk _samples were _analysed

for a..1-casein and 5355 for

₎

_{3-lactoglobulin.}

_a,,,-Cn

_c

_{homozygotes produced}

_{significantly}

more _a"1-casein than B

_homozygotes.

The /3_Lg

A

allele showed _{greater expression} both in

heterozygotes

and in _homozygotes than

_{the /3_Lg}

_B

allele. In

_{heterozygotes,}

the

_/3-Lg

_A

allele

produced nearly 50% more _{whey protein} than its

_{homologue. During}

the

_{spring-summer}

season

a"

1

-Cn

B

appeared

to _{synthesize more, relatively,} a,,-casein than

a"

1

-Cn°.

Possible

causes for this may be a _greater rate

_{of expression}

of the allele or increased

_{phosphorylation}

during spring-summer, producing

proportionally more a.,-casein.

cattle - milk _{protein genes - gene expression -} a"1-casein - /3-lactoglobulin

Résumé - _{Synthèse protéique} différentielle selon les variants de _{,6-lactoglobuline} et

de la caséine a"1 chez les bovins. Les _quantités de caséine a.1 et de

_{,6-lactoglobuline}

ont été déterminées dans le lait de 2059 vaches de race

Fleckvieh

et de race Braunvieh

de _Bavière; 6353 échantillons de lait ont été _analysés pour la caséine ce,1 et 5355 pour

la

_{,0-lactoglobuline.}

Les individus _homozygotes

aa

l

-Cn°

produisent significativement plus

de caséine que les individus homozygotes

.

D

-Cn

1

a.

L’expression de l’allèle

₍

_3-Lg

_A

est

supérieure à celle de l’allèle

₍

_3-Lg

_B

chez les individus _{hétérozygotes} ou _homozygotes. Chez les

_{hétérozygotes,}

l’allèle

_(3-LgA

a une _production de

protéine supérieure

d’environ 50%

à celle de son

_homologue.

Durant la _{période printemps-été,} l’allèle

0,1-Cn!

_synthétise

plus de caséine 0,1 que l’allèle

a.,i-Cn!.

Ceci pourrait provenir d’un taux d’expression

supérieur de l’allèle

as

l

- Cn

B

ou à une _augmentation de la

phosphorylation pendant

cette

période

produisant plus

de caséine a!0.

INTRODUCTION

In cattle rather few loci have been identified and efforts to link

_them

to

_quantitative

traits have not been _very successful. Milk

_protein

_genes,

_however,

are associated

with the

_quantitative

variation of the

_proteins

for which the codominant alleles are

coding. Moustgaard

et al.

_(1960),

Golikova and Panin

_(1972),

Michalak

_(1973),

Cer-bulis and Farrell

_(1975),

Komatsu et al.

_(1977),

Mariani et al.

_(1979),

McLean et al.

(1984),

Ng-Kwai-Hang

et al.

₍₁₉₈₇₎

and Aaltonen and Antila

₍₁₉₈₇₎

demonstrated

that the

_!3-Lg

_genotype AA

_produce

more

_{,0-lactoglobulin}

than _genotypes BB or

AB. Also McLean et al.

₍₁₉₈₄₎

on cattle and

_Boulanger

et al.

₍₁₉₈₄₎

and Grosclaude

et al.

₍₁₉₈₇₎

for _goats showed that

_a

_sl

_-Cn

_genotypes influence the

_production

of

a,,-casein.

Although

,!-Lg

and

_a

_sl

_-Cn

_genotypes show a different rate of

_protein

_synthesis,

there is little known about the

_expression

of the alleles in

_{heterozygotes.}

How-ever, the

haemoglobin

of sickle-cell

_heterozygote

is

_composed

of more than 60%

haemoglobin

A and less than 40% of

_haemoglobin

S

_(Wellis

and

Itano, 1951;

Wright-stone and

_Huisman,

_1968).

Such different rates of

_expression

_{of globin}

genes appear to be even more marked in Hb-C

_{heterozygotes}

(Boyer

et

_al.,

_1963;

_Itano,

₁₉₆₅₎

and

in thallasemias

_(Na-Nakorn

and

_{Wasi, 1970;}

Huisman et

_al.,

_1972).

Here we _report on differences in the concentration of

_a,,-caseins

and

_{#-lactoglobulins}

coded

_by

the

different alleles of

_{heterozygotes}

and

_homozygotes

of the Bavarian Simmental and

Bavarian Brown

_Alpine

cattle.

MATERIALS AND METHODS

The data are based on casein _resp.

_whey

_protein

_analysis

of 6353 _resp. 5355 milk

samples

from 2059 Simmental and 1809 Brown

_Alpine

cows. Simrrzental cows were

sampled

twice,

Brown

_Alpine

cows once. The statistical

_analysis

of Simmental

data was based on a model with effects of

_herd,

_{year-season, stage} and number of lactation and cows; that of the Brown

Alpine herd,

year-season,

stage

and number of

_lactation,

sire of the cow and _genotypes at 3 loci

_(in

the case of the

a,,-Cn

expression,

the

₃

_-Cn,

x-Cn and

_,0-Lg

_locus;

in the case of the

3

-Lg

expression,

the

a

si

-Cn, /!-Cn

and x-Cn

_locus).

The different mean

_expression

of the alleles

of

_heterozygous

_genotypes was tested

_by

a

_simple

_t-test; those of the

_homozygous

genotypes

by

the Student-Newman-Keuls test.

In Sirrcmental cows 2

_samples

were

_analysed

from

_nearly

_every cow. This

permitted

estimation of the

_{repeatability}

of the ratio of the

_proteins

in the

heterozygotes

(a

sl

-Cn

B

/a

s1

-Cn

c

resp.

!3-LgA/,Q-LgB).

The milk

_protein

content was measured

_by

the amido-black

method,

the

pro-portion

of the

_a

_si

_-casein

B resp. C and

0

-lactoglobulin

A resp. B

by

quantitative

photometric

determination from

_{cellogel electropherograms}

_(Kirchmeier,

_1975;

per-sonal

_{communication,}

_1988),

where the

_{optical density}

of the bands was measured

by

a

_{photodensitometer.}

The area under the

respective

peaks

was recorded and the

integral

area

_computed.

This

_corresponds

to the relative

_quantity

of the

_protein,

provided

that the

_{specific affinity}

to bind the

_dye

is taken into consideration.

!3-lactoglobulin

was isolated from

_whey

_proteins

after removal of a-lactalbumin

was achieved

_by

_{chromatofocusing (Sluyterman}

and

_Wijdenes,

_1978).

_Purity

and

homogeneity

was checked

_{by Page electrophoresis}

_(Raymond

and

_Weintraub,

_1959).

For determination of the

_{specific dye binding affinity,}

known

_quantities

of

!3-lactoglobulins

were

_{electrophorized,}

the bands coloured

_by

amido-black and measured

_{densitometrically.}

In

_comparison

with the standard

_{!3-lactoglobulin}

_A,

,Q-lactoglobulin

B had a

dye-binding

_activity

of

_1.05,

similar to

_published

results

(Reimerdes

and

_Mehrens,

_{1978; Krause,}

_personal

_{communication,}

_1988).

The

analogous

coefficient for _asl-casein B relative to asl-casein C was taken as

_1.06,

as

published previously by

McLean et al.

_(1982).

RESULTS

The _average differences between the

_expression

of

_a

_sl

_-casein

B and C alleles in

heterozygotes

were

_{insignificant}

(Table I).

_However,

_homozygous

a,,-Cn

cc

cows

had a

_higher

_asl-casein content than the alternative BB

_homozygote.

As shown in

Figure

1, the

_degree

of

_activity

of the alleles in the

_heterozygote

varied

_considerably

and its distribution

_approached

that of a normal curve.

The two alleles of

_{!3-lactoglobulin}

_heterozygote

_{,B_LgAB}

differed

_{significantly}

in

their

_activity.

₃

_-Lg

_A

_produced

about 50% more

_{lactoglobulin}

A than

/3-Lg

B

did

lactoglobulin

B. This difference is

_{paralleled by}

the difference between alternative

homozygotes.

The distribution

_{(Fig. 1)}

indicates considerable

_variability

and a

leptocurtosis.

In

_Figs.

2 to

_4,

the course over seasons in 2 _years of the ratio between the

_proteins

produced

by

the alleles of the

_{respective a,,-Cn}

and

₃

_{-Lg heterozygotes}

and the

expression

of the alleles in

_homozygotes

is shown. The difference between the

_whey

proteins

of the

_#-Lg

_{heterozygotes}

remains

_nearly

stable

_during

the _{2 years} of the

significantly

more

_synthetic

_activity

_during

the

_spring-

summer seasons than the

C-allele

_{(Fig. 2).}

Even in

_homozygous

_genotypes,

_a

_Sl

_-Cn

_B

shows more

_activity

in this

period

(Fig. 3).

In

_general,

_!i-LgB

and

a

sl

-Cn

C

show a more constant

_expression

in

_heterozygous

_genotypes than the resp.

homologous

alleles.

For the ratio of _Qs1-caseins in

_{heterozygotes, repeatability}

was estimated as

18%,

and as about 50% for the

_{,Q-lactoglobulins.}

This indicates that this ratio reflects to

a considerable

_degree

an innate _property of cows which

_probably

is inherited to a

large

extent.

However,

even for

whey

_protein,

a

_large

_proportion

of the

_variability

is due to factors not accounted for in the model. The lower

_{repeatability}

of the ratio between the _{caseins may} reflect inter alia the interaction between the allelic

activity

and seasonal influences. DISCUSSION

The two breeds Bavarian Simmental and Bavarian Brown

_Alpine

are located in

different

_regions

and the

_analysis

of the milk

_samples

was

_performed

at different

are the distributions and the seasonal

_changes.

As to seasonal effects on the ratio

of

caseins,

we can

_{only speculate}

at this time.

_{During spring-summer}

seasons, cows are either on _pasture or

_zero-grazing

and receive fresh _{grass which contains} steroids

The _{above average}

_expression

of the B allele in

a

sl

-Cn

heterozygotes

could,

to

some

degree,

be a

_product

of _aso-caseins of

_C-a

_s1

_protein

_co-migrating

with the

B-a

sl

protein.

For the B

_protein,

the contribution of the

a

so

-casein

is evident in the

_{electrophoregram}

and has been considered in

_estimating

the B-fraction. Also

the area in the case of

_homozygotes

was corrected for where indeed CC _genotypes

produce

significantly

more casein than the BB _genotypes.

_Therefore,

the above average

expression

of the B alleles in

heterozygotes during spring-summer

could be influenced also

_by

differences in

_{phosphokinase}

_{activity. However,}

the

_significant

ACKNOWLEDGEMENT

The authors are

_grateful

to the

referees

for constructive criticism. The

_{investigation}

was

_{supported by}

the Deutsche

_{Forschungsgemeinschaft.}

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