It　has　become　a　hot　topic　to　find　natural　food　substances　which　can　effectively　promote　calcium　absorption　in　human　body.　Given　this,　protein　hydrolysate　with　calcium-binding　capacity　was　prepared　from　fish　scales　(FSPH)　and　its　promotion　in　calcium　cellular　uptake　was　investigated　in　this　study.　The　binding　of　calcium　to　the　amino　nitrogen　atoms　and　the　carbonyl　oxygen　atoms　of　FSPH　led　to　the　structural　folding　and　aggregation.　The　calcium　cellular　uptake　efficiency　in　Caco-2　cells　population　model　was　significantly　increased　in　the　form　of　FSPH-calcium　complex.　Furthermore,　FSPH-Ca　could　effectively　resist　the　inhibitory　effect　of　dietary　inhibitors　on　calcium　cellular　uptake.　The　mRNA　expression　of　transient　receptor　potential　vanilloid　6　(TRPV6)　in　Caco-2　cells　was　remarkably　elevated　with　FSPH-Ca　treatment.　The　results　improved　the　understanding　of　the　interaction　mode　of　protein　hydrolysate　and　calcium,　and　provided　a　basis　for　developing　FSPH-Ca　as　functional　foods　to　improve　calcium　absorption.