Basal cell adhesion molecule/lutheran protein. The receptor critical for sickle cell adhesion to laminin.

Udani, Manisha; Zen, Qin; Cottman, Maisha J.; Leonard, Nicole; Jefferson, Shawn; Daymont, Carrie; Truskey, George A.; Telen, Marilyn J.

doi:10.1172/jci1204

Cited by 192 publications

(185 citation statements)

References 37 publications

(18 reference statements)

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“…HD SS RBC adhesion to laminin was completely blocked by srLU, while the CSS for static adhesion to laminin preincubated with BSA was 5.6 dyne/cm 2 for the specific blood sample shown, a result typical of the values we usually obtain when no exogenous protein is added (refer to Fig. 2) [15]. srLU also completely inhibited static adhesion of LD SS RBC to laminin (data not shown).…”

Section: Ss Rbc Static Adhesion To Laminin Is Mediated By B-cam/lusupporting

confidence: 64%

“…Packed RBC were diluted in phosphate-buffered saline (PBS) with 1% bovine serum albumin (BSA) and incubated with 3 mg/ml human laminin (Invitrogen), saturating dilutions of anti-Lu b mAb or a nonreactive control murine myeloma protein P3X63/Ag8 (P3) for 30 min, as previously described [15]. For laminin binding, cells were incubated at 37 C, instead of at room temperature, for 60 min, followed by washes in PBS and incubation with rabbit anti-laminin Ab and PE-conjugated second Ab.…”

Section: Flow Cytometrymentioning

confidence: 99%

“…Restriction mapping and DNA sequencing confirmed that the vector contained the desired cDNA sequence. The cDNA constructs were stably transfected into MEL cells by electroporation as previously described [15].…”

Section: Construction and Expression Of Lu Recombinant Proteinmentioning

confidence: 99%

“…Studies of RBC adhesion to immobilized laminin were performed using a variable-height flow chamber system as described previously [15]. In brief, clean glass slides were coated with 3 mg/ml laminin overnight at 4 C and were mounted into a variable-height flow chamber.…”

Section: Flow Chamber Assaysmentioning

confidence: 99%

“…The red cell laminin receptor B-CAM/LU [15] is the protein that bears Lutheran blood group antigens and binds specifically to laminins containing the a5 chain (laminins 10 and 11) [16,17]. B-CAM/LU comprises two protein spliceoforms derived from a single gene; B-CAM lacks the last 40 cytosolic amino acids of the LU protein [18,19].…”

Section: Introductionmentioning

confidence: 99%

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B‐CAM/LU expression and the role of B‐CAM/LU activation in binding of low‐ and high‐density red cells to laminin in sickle cell disease

et al. 2004

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Red blood cells from patients with sickle cell disease (SS RBC) adhere to laminin and over-express the high-affinity laminin receptor basal cell adhesion molecule/Lutheran protein (B-CAM/LU). This receptor has recently been shown to undergo activation in vitro through a protein kinase A-dependent mechanism. Low-density SS RBC express twothirds more B-CAM/LU than high-density SS RBC. However, high-density SS RBC have been identified as most adherent to laminin under flow conditions. We investigated the ability of low-and high-density SS RBC to interact with laminin under various conditions and explored factors that might be responsible for the differences in B-CAM/LU-laminin interaction between high-and low-density SS RBC. We confirmed that high-density SS RBC adhere to laminin more strongly than low-density SS RBC under flow conditions. However, low-density SS RBC bind soluble laminin most strongly and are the most adherent to laminin under static conditions. Soluble recombinant Lutheran extracellular domain protein completely blocked SS RBC adhesion to laminin under both static and flow conditions. The protein kinase A inhibitor 14-22 amide inhibited adhesion to laminin during flow by high-density SS RBC from patients with strongly adherent cells but had no effect on adhesion observed after a static phase. Deletion of the cytoplasmic domain of B-CAM as well as mutation of the juxtamembranous tyrosine residue failed to reduce B-CAM-mediated adhesion to laminin by transfected MEL cells. These studies confirm that B-CAM/LU is the most critical receptor mediating adhesion to laminin under both static and flow conditions. Dense SS RBC are most adherent to laminin despite bearing fewer laminin receptors, apparently due to a reversible protein kinase A-dependent process that is unlikely to involve direct phosphorylation of B-CAM/LU. Our results also suggest that the nature of the interaction of B-CAM/LU with laminin may be different under static and flow conditions. Am.

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Section: Ss Rbc Static Adhesion To Laminin Is Mediated By B-cam/lusupporting

confidence: 64%

Section: Flow Cytometrymentioning

confidence: 99%

Section: Construction and Expression Of Lu Recombinant Proteinmentioning

confidence: 99%

Section: Flow Chamber Assaysmentioning

confidence: 99%