Lactadherin (formerly BA46), a membrane-associated glycoprotein expressed in human milk and breast carcinomas, promotes Arg-Gly-Asp (RGD)-dependent cell adhesion.
Lactadherin, a major glycoprotein of the human milk fat globule membrane, is abundant in human breast milk and expressed in human breast carcinomas. Previously, we have shown that the mature protein, formerly known as BA46, has three domains: an epidermal growth factor (EGF)-like domain containing an Arg-Gly-Asp (RGD) cell adhesion sequence and C1 and C2 domains similar to those found in coagulation factors V and VIII. An alignment of lactadherin with its bovine (MGP57/53) and murine (MFG-E8) homologs shows that the RGD sequence has been conserved during evolution, suggesting that the RGD sequence is not fortuitous. We demonstrate that lactadherin purified using Triton X-114 phase partitioning promotes RGD-dependent cell attachment of green monkey kidney cells (MA104), mouse fibroblast cells (3T3-L1), and breast carcinoma cells (ELL-G). A lactadherin-specific monoclonal antibody, Mc3, inhibits attachment to purified lactadherin, suggesting that contaminants in the purification are not responsible for binding. In addition, the anti-integrin alpha(v)beta3 monoclonal antibody LM609 inhibits cell attachment of MA104 cells to lactadherin. These results demonstrate that lactadherin promotes RGD-dependent cell adhesion via integrins. Denaturation of lactadherin with heat and reducing conditions diminished cell attachment, suggesting that optimal cell attachment to RGD is dependent on the structural presentation of the sequence.
Taylor
M R
MR
Cancer Research Fund of Contra Costa, Walnut Creek, CA 94596, USA.
Couto
J R
JR
Scallan
C D
CD
Ceriani
R L
RL
Peterson
J A
JA
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D001943:Breast Neoplasms / Q000737:chemistry* / Q000473:pathology; D002277:Carcinoma / Q000737:chemistry* / Q000473:pathology; D002448:Cell Adhesion / Q000502:physiology*; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D002522:Chlorocebus aethiops; D017124:Conserved Sequence; D006801:Humans; D007668:Kidney; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D051379:Mice; D008894:Milk Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008895:Milk, Human / Q000737:chemistry*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism*; D011489:Protein Denaturation; D019030:Receptors, Vitronectin / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; C098942:MFGE8 protein, human; D008562:Membrane Glycoproteins; C490919:Mfge8 protein, mouse; D008894:Milk Proteins; D009842:Oligopeptides; D019030:Receptors, Vitronectin; C047981:arginyl-glycyl-aspartic acid