Limited proteolysis of the IGF binding protein-2 (IGFBP-2) by a specific serine protease activity in early breast milk.
IGF in milk possibly promote maturation of the gastrointestinal tract in newborns. We studied the composition of milk samples derived from 99 healthy women at regular intervals during a period beginning 3 d and ending 6 mo after birth. The concentrations measured by RIA on d 3 were 10.7+/-0.4 ng/mL for IGF-II, 1.9+/-0.1 ng/mL for IGF-I, 100+/-5 ng/mL for IGF binding protein-3 (IGFBP-3), and 2163+/-108 ng/mL for IGFBP-2. All factor concentrations decreased by up to 70% in the course of the 6 mo. The most striking finding was an IGFBP-2-specific protease activity. Protease assays performed by incubation of 125I-IGFBP-2 with milk yielded fragments of 14, 16, 23, and 25 kD. 125I-IGFBP-3 was not cleaved. Proteolysis occurred only in milk from mothers until 4 wk postpartum and could be visualized by immunoblots. Since the affinity of the fragments to 125I-IGF-II was low, they were not demonstrable by ligand blot. Inhibitor studies and pH optimizing classified the IGFBP-2 protease as an Me2(+)-dependent serine protease with a pH optimum of 7 to 8. The proteolytic activity of further milk proteins, known as IGFBP proteases, was analyzed. Epidermal growth factor receptor peptide and prostate-specific antigen did not cleave IGFBP-2, although the protease activity correlated (r = 0.84, p < 0.00003) with the prostate-specific antigen concentration in milk. The y-nerve growth factor cleaved 125I-IGFBP-2, but in a completely different manner than the milk protease. In conclusion, the IGFBP-2 protease in milk is most probably a kallikrein. The specific proteolysis of IGF/IGFBP-2 complexes may increase the biologic availability of IGF in early milk. This mechanism may promote growth of the maternal breast epithelium and maturation of the gastrointestinal tract of newborns.
Neonatology Department, University Children's Hospital, Tuebingen, Germany.
M W
MW
Elmlinger
R
R
Grund
M
M
Buck
H A
HA
Wollmann
N
N
Feist
M M
MM
Weber
C P
CP
Speer
M B
MB
Ranke
D005260:Female; D006801:Humans; D018971:Insulin-Like Growth Factor Binding Protein 2 / Q000378:metabolism*; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000378:metabolism; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D007335:Insulin-Like Growth Factor II / Q000378:metabolism; D007700:Kinetics; D007774:Lactation; D008895:Milk, Human / Q000737:chemistry / Q000201:enzymology*; D012044:Regression Analysis; D012697:Serine Endopeptidases / Q000378:metabolism*; D013379:Substrate Specificity; D013997:Time Factors
D018971:Insulin-Like Growth Factor Binding Protein 2; D018972:Insulin-Like Growth Factor Binding Protein 3; D007334:Insulin-Like Growth Factor I; D007335:Insulin-Like Growth Factor II; D012697:Serine Endopeptidases